Stem Cell-Based Regenerative Medicine in Canine Practice E-Book

Mudasir Bashir Gugjoo

• Herausgeber: John Wiley & Sons
• Kategorie: Fachliteratur
• Sprache: Englisch

Seitenzahl: 1134

Veröffentlichungsjahr: 2025

Beschreibung

A practical guide to stem cell-based regenerative techniques in dogs

Stem Cell-Based Regenerative Medicine in Canine Practice delivers an essential reference for the use of stem cells in canine practice. Intuitively organized by organ and body systems, the book is easy-to-follow, and addresses many common conditions encountered when dealing with dogs in veterinary practice, including those affecting the central nervous system, muscular system, and vascular system.

The book is grounded in evidence-based research and readers are provided with guidance on the potential challenges and problems that may arise when considering and using stem cell therapeutics. It also explains the techniques for stem cell extraction and how to use stem cell regenerative medicine for the best results.

Readers will also find:

• An introduction to the treatment of various clinical conditions like corneal ulcers and keratoconjunctivitis
• Comprehensive explorations of the treatment of osteoarthritis, bone gap defects, tendon/ligament injuries, muscle affections, peripheral nerves, periodontitis, esophageal ulcers, inflammatory bowel disease, liver, diabetes, and skin wounds
• Practical discussions of canine mesenchymal stem cell sources, properties, and regulations for clinical use
• An overview of comparative analysis between various stem cells for therapeutics
• Complimentary access to a website containing reference videos and clinical guidance

Written for veterinarians in general practice, Stem Cell-Based Regenerative Medicine in Canine Practice will also benefit veterinary students and researchers.

Leseprobe

Table of Contents

Cover

Table of Contents

Title Page

Copyright Page

About the Author

Foreword

Preface

Abbreviations

About the Companion Website

1 Canine Mesenchymal Stem Cells Sources, Properties, and Regulations for Clinical Use

1.1 Introduction

1.2 Understanding the Stem Cell

1.3 Sources of Dog MSCs for Proliferation

1.4 Why Do We Need to Culture Mesenchymal Stem Cells?

1.5 Why to Characterize MSCs?

1.6 Factors Affecting MSCs Therapeutic Outcome

1.7 Stem Cell Regulations in Research and Therapeutics

References

2 Osteoarthritis

2.1 Introduction

2.2 Predisposing Factors

2.3 OA Pathogenesis

2.4 MSCs and Their Potential Therapeutic Role in OA

2.5

In Vivo

MSCs Studies

References

3 MSCs in Canine Bone Tissue Engineering

3.1 Introduction

3.2 Mesenchymal Stem Cells and Bone Regeneration

3.3

In Vivo

Studies

References

4 Muscle, Tendon, and Ligament Affections

4.1 Introduction

4.2 Why Mesenchymal Stem Cells and Not Tendon Stem/Progenitor Cells (TSPCs)?

4.3

In Vivo

Studies

4.4 Muscle Studies

References

5 Peripheral Nerve Affections

5.1 Introduction

5.2 Peripheral Nerve Injury

5.3 Why MSCs and Not Schwann Cells for Nerve Regeneration

5.4

In Vivo

Peripheral Nerve Injury Studies

References

6 Central Nervous System Affections

6.1 Introduction

6.2 CNS Intrinsic Regeneration Potential

6.3 Exogenous Stem Cell Therapy

References

7 Myocardial and Valvular Diseases

7.1 Introduction

7.2 Why MSCs in Cardiac Diseases?

7.3

In Vivo

MSCs Therapeutic Studies in Dogs

References

8 Oral and Esophageal Affections

8.1 Introduction

8.2 Oral Mucosal Ulcers

8.3 Periodontal Disease

8.4 Maxillary Alveolar Bone Cleft Model

8.5 Peri‐implant Defect Models

8.6 Esophagus

References

9 Inflammatory Bowel Disease and Anal Furunculosis

9.1 Introduction

9.2 Why Mesenchymal Stem Cells

9.3

In Vivo

Studies

References

10 Liver Affections

10.1 Introduction

10.2 Regenerative Medicine in Canine Hepatology

10.3

In Vivo

Liver Regenerative Studies

10.4 Hepatocutaneous Syndrome

References

11 Lung Affections

11.1 Introduction

11.2 MSCs Potential Therapeutic Effects

11.3

In Vivo

Studies

References

12 Renal Insufficiency and Urinary Bladder Affections

12.1 Introduction

12.2

In Vitro

Renal Studies

12.3

In Vivo

Renal Studies

12.4 Urinary Bladder

References

13 Skin Wounds and Atopic Dermatitis

13.1 Introduction

13.2

In Vivo

Skin Wound‐Healing Studies Using MSCs

13.3 Atopic Dermatitis (AD)

References

14 Diabetes

14.1 Introduction

14.2

In Vitro

Studies

14.3

In Vivo

Studies

References

15 Corneal Ulcers and Keratoconjunctivitis Sicca

15.1 Introduction

15.2 Mesenchymal Stem Cells (MSCs) Versus Corneal/Limbal Stem Cells

15.3 Corneal Affections

15.4 Keratoconjunctivitis Sicca (KCS)

References

Index

End User License Agreement

List of Tables

Chapter 2

Table 2.1 Dog preclinical experimental chondral defect models evaluating me...

Table 2.2 Dog preclinical experimental femoral osteochondral defect and tro...

Table 2.3 Dog

in vivo

MSCs preclinical experimental studies on joint tissue...

Table 2.4 Clinical studies on evaluation of mesenchymal stem cells therapeu...

Table 2.5 Clinical studies on evaluation of MSCs therapeutic effect on oste...

Table 2.6 Clinical studies on evaluation of MSCs therapeutic effect on oste...

Table 2.7 Clinical studies on evaluation of MSCs therapeutic effect on dysp...

Chapter 3

Table 3.1 Experimental studies on femoral bone defect in dogs using mesench...

Table 3.2 Experimental studies on radial and ulnar bone defect in dogs util...

Table 3.3 Experimental studies on tibial bone defects in dogs using mesench...

Table 3.4 Experimental studies on healing of mandibular bone defects and cl...

Table 3.5 Dog

in vivo

clinical studies on various bone affections using mes...

Chapter 4

Table 4.1 Dog

in vivo

preclinical experimental studies on flexor tendon mod...

Table 4.2 Dog

in vivo

preclinical experimental studies on dog infraspinatus...

Table 4.3 Dog

in vivo

MSCs clinical studies on tendinopathy of gastrocnemiu...

Table 4.4 Dog

in vivo

clinical studies on cranial cruciate ligament rupture...

Table 4.5 Dog

in vivo

clinical studies on muscle affections evaluating heal...

Chapter 5

Table 5.1 Preclinical experimental studies on regeneration of dog sciatic n...

Table 5.2 Preclinical experimental studies on regeneration of resected dog ...

Table 5.3 Preclinical experimental study on regeneration of ulnar nerve def...

Chapter 6

Table 6.1 Dog clinical studies on canine distemper evaluating the therapeut...

Table 6.2 Dog preclinical experimental studies evaluating mesenchymal stem ...

Table 6.3 Dog preclinical experimental studies evaluating mesenchymal stem ...

Table 6.4 Clinical studies on spinal cord injuries (SCIs) in dogs evaluatin...

Table 6.5 Dog Clinical intervertebral disc disease (IVDD)/herniation studie...

Chapter 7

Table 7.1 MSCs preclinical experimental studies on dog myocardial infarctio...

Table 7.2 MSCs preclinical experimental studies on biological pacemakers in...

Table 7.3 MSCs clinical studies on cardiomyopathy and valvular diseases in ...

Chapter 8

Table 8.1 Preclinical experimental studies on induced oral mucosal ulcers i...

Table 8.2 Preclinical experimental studies in periodontal disease involving...

Table 8.3 Preclinical experimental studies on furcation defects in dogs

Table 8.4 Preclinical experimental studies in periodontal disease involving...

Table 8.5 Preclinical experimental studies on esophageal mucosal resection ...

Chapter 9

Table 9.1 Clinical studies on inflammatory bowel disease and anal furunculo...

Chapter 10

Table 10.1 MSCs preclinical experimental studies on acute and chronic liver ...

Chapter 11

Table 11.1 Preclinical experimental studies on dog cardiopulmonary bypass m...

Chapter 12

Table 12.1 MSCs preclinical experimental studies on renal injury and bladde...

Table 12.2 Clinical studies on chronic kidney disease and Fanconi syndrome ...

Table 12.3 Urinary bladder resections evaluating therapeutic role of mesenc...

Chapter 13

Table 13.1 Preclinical experimental studies on full‐thickness skin wounds i...

Table 13.2 Clinical studies on skin wounds in dog evaluating mesenchymal st...

Table 13.3 Preclinical and clinical studies on atopic dermatitis in dog eva...

Chapter 14

Table 14.1 Preclinical experimental studies on diabetes in dog using evalua...

Table 14.2 MSCs clinical studies on insulin‐dependent diabetes mellitus in ...

Chapter 15

Table 15.1 Clinical studies of dog corneal affections treated with mesenchy...

Table 15.2 Clinical studies on keratoconjunctivitis in dogs treated with me...

List of Illustrations

Chapter 1

Fig. 1.1 Sources of stem cells from initial fertilization to the adulthood o...

Fig. 1.2 Sources of mesenchymal stem cells and their characterization on the...

Chapter 2

Fig. 2.1 Predisposing factors of osteoarthritis in dogs.

Fig. 2.2 Osteoarthritis in dogs mediated through cartilage: role of inflamma...

Fig. 2.3 Osteoarthritis in dogs mediated through cartilage: role of growth f...

Fig. 2.4 Osteoarthritis in dogs mediated through cartilage: role of receptor...

Fig. 2.5 Osteoarthritis in dogs mediated through synovitis: role of inflamma...

Fig. 2.6 Osteoarthritis in dogs mediated through subchondral bone: role of e...

Fig. 2.7 Factors affecting mesenchymal stem cells chondrogenic potential.

Fig. 2.8 Therapeutic mechanisms that mesenchymal stem cells may offer in ost...

Fig. 2.9 Percentage of different experimental osteoarthritis models conducte...

Fig. 2.10 Percentage of different clinical osteoarthritis studies in dogs ev...

Chapter 3

Fig. 3.1

Ex vivo

osteogenic differentiation of mesenchymal dog stem cells th...

Fig. 3.2

In vivo

bone healing potential of mesenchymal stem cells in differe...

Fig. 3.3

In vivo

osteogenic healing potential of mesenchymal stem cells in d...

Chapter 4

Fig. 4.1 Three‐stage process of affection in tendon and ligament: reactive, ...

Fig. 4.2 Comparative analysis of stem/progenitor cells of tendon or ligament...

Fig. 4.3 Growth factors and their effect on mesenchymal stem cell tenogenesi...

Fig. 4.4 (A) Differentiation of mesenchymal stem cells into tenocyte‐like ce...

Fig. 4.5 Percentage of different tendinopathy models in dogs that are evalua...

Fig. 4.6 Number of clinical studies utilizing mesenchymal stem cells in tend...

Fig. 4.7 Number of clinical studies utilizing mesenchymal stem cells in trau...

Chapter 5

Fig. 5.1 Neuron and its different parts including soma (cell body), axon, an...

Fig. 5.2 Schematic representation of different grades of nerve injuries as p...

Fig. 5.3 Nerve regeneration in low‐grade peripheral nerve injuries involving...

Fig. 5.4 Role of Schwann cells in peripheral nerve regeneration.

Fig. 5.5 Pie chart showing different peripheral nerve injury models conducte...

Chapter 6

Fig. 6.1 Schematic diagram showing

ex vivo

differentiation of mesenchymal st...

Fig. 6.2 Potential therapeutic effects that mesenchymal stem cells may impar...

Fig. 6.3 Pie chart shows two types of stem cells (MSCs and OESCs) that are u...

Fig. 6.4 The pie chart shows different types of canine spinal cord injury mo...

Fig. 6.5 The pie chart shows mesenchymal stem cells are clinically evaluated...

Chapter 7

Fig. 7.1 Mesenchymal stem cells therapeutic properties:

ex vivo

differentiat...

Fig. 7.2 Experimental cardiac regeneration models conducted in dogs using me...

Fig. 7.3 Clinical trials for different cardiac ailments conducted in dogs us...

Chapter 8

Fig. 8.1 List of different oral affections in dogs that are evaluated for th...

Fig. 8.2 Pie chart showing a number of studies conducted on a particular ora...

Fig. 8.3 Pie chart showing esophageal affections and the number of studies c...

Chapter 9

Fig. 9.1 Different forms of chronic enteropathies in dogs. Inflammatory bowe...

Fig. 9.2 (A) It shows the co‐ordination of the epithelial mucosal barrier, i...

Fig. 9.3 Anal furunculosis pathogenicity and features in dogs.

Fig. 9.4 The figure demonstrates why mesenchymal stem cells are preferred fo...

Fig. 9.5 Number of studies conducted in dogs that evaluate mesenchymal stem ...

Chapter 10

Fig. 10.1 Schematic diagram showing different manipulation techniques employ...

Fig. 10.2 Experimental studies conducted in dogs evaluating MSCs’ therapeuti...

Chapter 11

Fig. 11.1 Effect of microenvironment on mesenchymal stem cell properties and...

Fig. 11.2 Potential therapeutic effects that the rejuvenated or extra‐lung m...

Chapter 12

Fig. 12.1 Two different regenerative medicine methods undertaken for renal a...

Fig. 12.2 Internal repair potential in kidney is attributed either to dediff...

Fig. 12.3 Four different

ex vivo

methods to differentiate mesenchymal stem c...

Fig. 12.4 Pie chart showing number of different renal models that evaluate m...

Chapter 13

Fig. 13.1 Pic chart showing number of clinical full‐thickness skin wound mod...

Fig. 13.2 Pic chart showing number of experimental and clinical studies on a...

Chapter 14

Fig. 14.1 Diabetes‐related data in canine population. Around 1.2% prevalence...

Fig. 14.2

Ex vivo

differentiation of mesenchymal stem cells into insulin‐pro...

Fig. 14.3 Pie chart showing

in vivo

experimental and clinical studies on dia...

Chapter 15

Fig. 15.1 Comparative analysis of corneal/limbal stem cells and the mesenchy...

Fig. 15.2 Potential therapeutic effects that may be offered by mesenchymal s...

Fig. 15.3 Pie chart showing various, although very limited, corneal affectio...

Fig. 15.4 Pie chart showing the number of keratoconjunctivitis sicca clinica...

Guide

Cover Page

Table of Contents

Title Page

Copyright Page

About the Author

Foreword

Preface

Abbreviations

About the Companion Website

Begin Reading

Index

WILEY END USER LICENSE AGREEMENT

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Stem Cell‐Based Regenerative Medicine in Canine Practice

Mudasir Bashir Gugjoo

Division of Veterinary Clinical ComplexFaculty of Veterinary Sciences & Animal HusbandrySher-e-KashmirUniversity of Agricultural Sciences and Technologyof KashmirShuhama, Alusteng, Srinagar, Jammu and KashmirIndia

This edition first published 2025© 2025 John Wiley & Sons Ltd

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Library of Congress Cataloging‐in‐Publication DataNames: Gugjoo, Mudasir Bashir, author.Title: Stem cell‐based regenerative medicine in canine practice / Mudasir  Bashir Gugjoo.Description: Hoboken, NJ : Wiley, 2025. | Includes bibliographical  references and index.Identifiers: LCCN 2024058459 (print) | LCCN 2024058460 (ebook) | ISBN  9781394253258 (hardback) | ISBN 9781394253272 (adobe pdf) | ISBN  9781394253265 (epub)Subjects: MESH: Dog Diseases–therapy | Stem Cell  Transplantation–veterinary | Tissue Engineering–veterinary |  Regenerative Medicine–methodsClassification: LCC SF991 (print) | LCC SF991 (ebook) | NLM SF 991 | DDC  636.70896–dc23/eng/20250213LC record available at https://lccn.loc.gov/2024058459LC ebook record available at https://lccn.loc.gov/2024058460

Cover Design: WileyCover Images: © Mudasir Bashir Gugjoo

About the Author

Dr. Mudasir Bashir Gugjoo, PhD, is a distinguished faculty member at Sher‐e‐Kashmir University of Agricultural Sciences & Technology of Kashmir (SKUAST‐K), specializing in Veterinary Surgery and Radiology. He completed his doctoral studies in Veterinary Surgery and Radiology at the Indian Veterinary Research Institute (IVRI), Izatnagar, focusing on stem cell‐based therapeutics. Dr. Gugjoo has been widely recognized for his contributions to veterinary science, receiving multiple accolades, including awards for best paper presentations and faculty teaching. His academic journey was supported by prestigious fellowships, such as the Junior and Senior Research Fellowships from the Indian Council for Agricultural Research (ICAR), funding his master’s and doctoral studies.

Currently, Dr. Gugjoo's research is funded by the Science and Engineering Research Board (SERB) and the Department of Biotechnology (DBT), Government of India (GoI). His laboratory focuses on exploring the regenerative potential of mesenchymal stem cells and developing innovative tissue‐engineered biomaterials aimed at regenerating bone, cartilage, wounds, peripheral nerves, and corneal ulcers. He has successfully mentored numerous master's and doctoral students in regenerative medicine and organized several workshops and hands‐on training sessions, sponsored by SERB and the Indian Council of Medical Research (ICMR), GoI.

Dr. Gugjoo and his team have led pioneering advancements in regenerative medicine, developing tissue‐engineered constructs for cartilage, bone, corneal ulcers, and specialized skin creams. In veterinary cardiology, they have established standardized echocardiographic parameters and indices for different canine breeds. Additionally, his team has refined a sonographic technique to accurately diagnose specific brain parasites, advancing diagnostic capabilities in veterinary medicine.

Dr. Gugjoo has an extensive body of work, having authored over 70 peer‐reviewed publications and written and edited books on stem cell research in veterinary science with both national and international publishers. He has contributed chapters to various academic books and frequently writes columns in newspapers. In addition to his research and teaching, he has been an active member of scientific societies, participated in faculty improvement programs, and served as a resource person in conferences and workshops. His work has been cited in leading veterinary texts and journals, and he also contributes as an editor and reviewer for peer‐reviewed journals and research funding proposals.

Foreword

It is a pleasure to write a foreword for the book Stem Cell‐Based Regenerative Medicine in Canine Practice, authored by Mudasir Bashir Gugjoo. Veterinary medicine has witnessed remarkable advancements over the past few decades, with regenerative medicine emerging as a beacon of hope for treating various clinical conditions in our canine companions. This book delves into the transformative potential of mesenchymal stem cells (MSCs) in addressing a myriad of health issues that afflict dogs.

The author has meticulously compiled and presented comprehensive insights into the therapeutic mechanisms of MSCs, highlighting their ability to differentiate ex vivo and their therapeutic effects in vivo. This book covers a wide spectrum of clinical conditions affecting different tissues and organs, offering a detailed exploration of how MSCs can be harnessed to promote healing and regeneration.

From osteoarthritis to tendinopathies, from liver affections to neurological disorders, and from immune affections to infections, the potential applications of MSCs are vast and varied. The authors provide a thorough examination of both experimental and clinical studies, showcasing the promising results and the challenges that lie ahead in this rapidly evolving field. The book also puts to rest the media reports of hyped‐up claims of miraculous cures from stem cells that currently lack scientific backing.

As we stand on the cusp of a new era in veterinary medicine, this book serves as an invaluable resource for veterinarians, researchers, and students alike. It not only underscores the scientific advancements but also emphasizes the practical applications and future directions of stem cell‐based therapies in canine practice. Moreover, in numerous cases, dogs are used as model animals for human translational studies; the literature and treatments developed therefrom for humans can also become therapeutic options for dogs. This cross‐species relevance highlights the “One Health” principle, which emphasizes the interconnectedness of animal, human, and environmental health.

Dr. Mudasir Bashir Gugjoo, the author of this book, brings a wealth of knowledge and experience to the table. His dedication to advancing veterinary medicine and his pioneering work in the field of regenerative therapies have been instrumental in shaping this comprehensive guide.

I am confident that this book will inspire and guide not only those dedicated to improving the health and well‐being of companion pets but also the ones interested in stem cell biology. The journey of regenerative medicine is just beginning, and this book is a testament to the incredible potential that lies ahead.

Nazir Ahmad Ganai

Vice Chancellor, SKUAST‐K

Preface

In today’s world, pet owners are more determined than ever to provide the best possible care for their animals. While regenerative medicine is not yet a cure‐all, it holds great promise for alleviating some of the common clinical issues faced by canines. Stem cell‐based therapies, in particular, are generating interest for their potential to offer new solutions in veterinary medicine. However, the rapidly growing and diverse body of literature on this topic can make it challenging for veterinarians, researchers, and even pet owners to grasp the current state of regenerative medicine and its therapeutic possibilities.

This book aims to fill a significant gap by addressing the common clinical conditions in canines that could benefit from regenerative medicine, with a special focus on stem cell‐based therapies. It provides a thorough discussion of the problems and limitations in current treatment approaches and explores how regenerative medicine utilizing stem cells, with or without the use of biomaterials, could offer new hope in canine practice. The book reviews the existing literature on the therapeutic potential of stem cells in canines, bringing an evidence‐based perspective to the table.

While much of the available literature on regenerative medicine is focused on human medicine, there is a pressing need for a comprehensive guide dedicated to veterinary applications—especially for dogs. While the author’s previous works have covered stem cell therapeutics in veterinary medicine, this book is unique in its focus on canine clinical practice, making it an essential, one‐stop reference for anyone interested in the subject.

Designed primarily as a reference guide for veterinary practitioners, stem cell researchers, and students, this book will also serve as an informative resource for medical professionals and pet owners eager to understand the potential of stem cell‐based regenerative medicine. By highlighting the limitations in current treatments and the gap between laboratory research and practical applications, this book aims to inspire future research and advance the field of regenerative medicine for the benefit of canine health.

Abbreviations

18

F‐FDG:

F‐fluorodeoxyglucose

2D:

2‐Dimensional

3‐D:

3 dimensional

4HNE:

4‐Hydroxynenonal

5‐Aza:

5‐Aza‐2′‐deoxycytidine

5‐HT:

Serotonin

99m

Tc‐MIBI:

99m

Tc‐sestamibi

99mTcO4−:

99mTc‐pertechnetate

ABCG2:

ATP‐binding cassette (ABC) transporters

AC:

Adenylate cyclase

ACI:

Autologous chondrocyte implantation

ACM:

Acellular matrix

ACTA2:

Actin alpha 2

AD:

Adipose tissue

ADAMTS:

Disintegrin and metalloproteinase thrombospondin‐like motifs

ADAMTS16:

ADAM metallopeptidase with thrombospondin type 1 motif 16

AF:

Anal furunculosis

AFP:

Alpha‐fetoprotein

AKI:

Acute kidney injury

AKT:

Ak strain transforming

ALB:

Albumin

ALDH1A3:

Aldehyde dehydrogenase 1A3

ALI:

Acute lung injury

ALK5:

Activin receptor‐like kinase 5

ALP:

Alkaline phosphatase

ALT:

Alanine transaminase

AM:

Amniotic membrane

AMPs:

Antimicrobial peptides

ANG:

Angiopoietin

Anpep:

Aminopeptidase N

AP:

Apical ligament

AQP1:

Aquaporin 1

ARDS:

Acute respiratory distress syndrome

ARE:

Antibiotic responsive enteropathy

AST:

Aspartate aminotransferase

AT3:

Antithrombin 3

ATF6:

Activating transcription factor 6

ATMPs:

Advanced therapy medicinal products

ATP:

Adenosine triphosphate

ATR:

Angiotensin receptor

AURKA:

Aurora‐A kinase

AV:

Atrioventricular

Bad:

Bcl‐2–associated death protein

Bax:

Bcl‐2–associated X protein

BBB:

Blood–brain barrier

BCL‐2:

B‐cell lymphoma 2

BCP:

Biphasic calcium phosphate

BC‐TS:

Tubular bacterial cellulose

BDKRB2:

Bradykinin receptor B2

BDNF:

Brain‐derived nerve factor

BDNF:

Brain‐derived neurotrophic factor

bFGF:

Basic fibroblast growth factor

BglII:

Bacillus globigii‐derived type II restriction endonuclease

BHA:

Butylated hydroxyanisole

BIC:

Bone implant contact

BM‐:

Bone marrow

BMA:

Bone marrow aspirate

BMD:

Bone mineral density

BME:

β‐Mercaptoethanol

BMP:

Bone morphogenetic factor

BrdU:

Bromodeoxyuridine

BST1:

Bone marrow stromal cell antigen‐1

BTE:

Bone tissue engineering

BUN:

Blood urea nitrogen

C/ABB:

Chitosan/anorganic bovine bone

CADESI‐4:

Canine atopic dermatitis extent severity index‐4

CAGR:

Compound annual growth rate

cAMP:

3′5‐cyclic monophosphate monohydrate

CARD15:

Caspase recruitment domain‐containing protein 15

CBC:

Complete blood count

CBPI:

Canine Brief Pain Inventory

CCAAT/EBPα:

Cytosine–cytosine–adenosine–adenosine–thymidine enhancer binding protein

CCECAI:

Canine Chronic Enteropathy Clinical Activity Index

CCL2:

C‐C motif ligand 2

CCl

4

:

Carbon tetrachloride

CCNB1:

Cyclin B1

CD:

Canine distemper

CD:

Cluster of differentiation

CDCs:

Cardiosphere‐derived stem cells

CdHA:

Calcium‐deficient hydroxyapatite

CDK1:

Cyclin‐dependent kinase 1

Cdkn1a:

Cyclin‐dependent kinase inhibitor 1A

CDV:

Canine distemper virus

CDX4:

Caudal Type Homeobox 4

CFU(f):

Colony forming unit(fibroblasts)

CGRP:

Calcitonin gene‐related peptide

CHF:

Congestive heart failure

CHOP:

C/EBP Homologous Protein

CIBDAI:

Clinical Inflammatory Bowel Disease Activity Index

CIE:

Chronic inflammatory enteropathy

CIMVs:

Cytochalasin B‐induced membrane vesicles

C‐jun:

Jun Proto‐Oncogene

CK19:

Cytokeratin19

CKD:

Chronic kidney disease

CLEC3A:

C‐type lectin domain family 3 member A

CM:

Conditioned medium

CMAP:

Compound muscle action potential

CMC:

Carboxy methyl cellulose

CMR:

Cardiovascular magnetic resonance

c‐MYC:

Cellular myelocytomatosis

CNS:

Central nervous system

CNTF:

Ciliary neurotrophic factor

COL2A:

Collagen 2A

COL3A1:

Collagen 3A1

Coll I:

Collagen I

COX:

Cyclooxygenase

CPB:

Cardiopulmonary bypass

CPS:

Calcium phosphate scaffold

CrCL:

Cranial cruciate ligament

CRI:

Continuous rate infusion

CRISPR:

Clustered regularly interspaced short palindromic repeats

CRP:

C‐reactive protein

CSA:

Cyclosporin A

CSF:

Cerebrospinal fluid

CSG:

Chondroitin sulphate glycosaminoglycan

CSH:

Calcium sulfate hemihydrate

CTGF:

Connective tissue growth factor

CTI:

Cardiac troponin I

CTNND2:

Catenin delta 2

CTT:

Cardiac troponin T

CVM:

Centre for veterinary medicine

Cx45:

Connexin 45

CXCR4:

C‐X‐C chemokine receptor type 4

CYP:

Cytochrome P450

DAPT:

N‐[N‐(3,5‐Difluorophenacetyl)‐

L

‐alanyl]‐S‐phenylglycine t‐butyl ester

dbCAMP:

Dibutyryl Cyclic Adenosine Monophosphate

DCM:

Dilatation cardiomyopathy

DCN:

Decorin

DCs:

Dendritic cells

DDFT:

Deep digital flexor tendon

DE:

Definitive endoderm

DED:

Dry eye disease

Dexa:

Dexamethasone

DFE:

Dermatophagoides farinae

extract

DISA:

Dual isotope simultaneous acquisition

DKA:

Diabetic ketoacidosis

DKK1:

Dickkopf‐1

DLA:

Dog leukocyte antigen

DM:

Diabetes mellitus

DMD:

Duchenne’s muscle dystrophy

DMEM:

Dulbecco’s minimally essential medium

DMEM‐HG:

Dulbecco’s modified Eagle’s Media‐High Glucose

dMSCs:

Dog mesenchymal stem cells

DMSO:

Dimethyl sulfoxide

dMyHC:

Developmental myosin heavy chain

DNA:

Deoxyribonucleic acid

DP:

Dental pulp

DRB1:

Major Histocompatibility Complex, Class II, DR Beta 1

DTEF:

Divergent transcriptional enhancer factor

ECG:

Electrocardiography

ECM:

Extracellular matrix

ECMO:

Extracorporeal membrane oxygenation

EDNRA:

Endothelin receptors

EDVI:

End diastolic volume index

EF:

Ejection fraction

EGF:

Epidermal growth factor

EGFP:

Enhanced green fluorescent protein

Egr1:

Early response protein

ELISA:

Enzyme‐linked immunosorbent assay

EMT:

Epithelial–mesenchymal transition

eNOS:

Endothelial nitric oxide synthase

Eomes:

Eomesodermin

Epac2:

Exchange Protein Directly Activated by cAMP 2

EPCS:

Electric‐pulse current stimulation

EPCs:

Endothelial progenitor cells

EPO:

Erythropoietin

EPOR:

Erythropoietin receptor

ER:

Endoplasmic reticulum

ERK:

Extracellular signal‐regulated kinase

ESCs:

Embryonic stem cells

ESD:

Endoscopic submucosal dissection

ESF:

External skeletal fixation

ESVI:

End systolic volume index

EVs:

Extracellular vesicles

FBS:

Fetal bovine serum

FDA:

Food and Drug Administration

FDMBB:

Freeze‐dried mineral bone block

FGF:

Fibroblast growth factor

FGFR:

Fibroblast growth factor receptor 2

FGFR1:

Fibroblast growth factor 1 receptor

FISH:

Fluorescence

in situ

hybridization

Foxa1:

Forkhead box protein A genes

FOXP3:

Forkhead box protein P3

FRE:

Food responsive enteropathy

FS:

Fractional shortening

Fzd:

Frizzled

GAD65:

Glutamic acid decarboxylase 65

GADA:

Glutamic acid decarboxylase‐65 antibody

GAG:

Glycosaminoglycans

GalC:

Galactocerebrosidase

GAP43:

Growth‐associated protein 43

GAP‐43:

Growth‐associated protein 43

GATA:

Guanine Adenine Thiamine Adenine (GATA) binding protein 4

GBR:

Guided bone regeneration

GCPs:

Good clinical practices

GDF‐6:

Growth differentiation factor‐6

GDNF:

Glial cell line‐derived neurotrophic factor

GDNF:

Glial cell‐derived neurotrophic factor

GFAP:

Glial fibrillary acidic protein

GFR:

Glomerular filtration rate

GLP‐1:

Glucagon‐like peptide‐1

Glut2:

Glucose transporter 2

GLUT‐4:

Glucose transporter type 4

GM:

Gentamicin

GM‐CSF:

Granulocyte macrophage colony‐stimulating factor

GMPs:

Good manufacturing practices

GNL3:

Guanine nucleotide‐binding protein‐like 3

GSCs:

Gastrulation stage cells

GSH:

Reduced glutathione

GSK3β:

Glycogen Synthase Kinase 3β

GWS:

Genome‐wide association studies

HA:

Hydroxyapatite or hyaluronic acid as specified

HBDS:

Heparin/fibrin‐based delivery system

HCN4:

mouse hyperpolarization‐activated cyclic nucleotide‐gated cation

HCPI:

Helsinki Chronic Pain Index

HCS:

Hepatocutaneous syndrome

HD:

Heart disease

Hes1:

Hairy and enhancer of split‐1

hESCs:

human embryonic stem cells

HF:

Heart failure

HGF:

Hepatocyte growth factor

HindIII:

Type II site‐specific deoxyribonuclease restriction enzyme

hJ:

Human jaw

hMSCs:

Human mesenchymal stem cells

Hnf:

Hepatocyte nuclear factor

HNF4:

Hepatocyte nuclear factor‐4 alpha

HO‐1:

Hemoxygenase 1

Hoxb4:

Homeobox 4

HSP:

Heat shock protein

hTID:

Human type 1 DM

HUVECs:

Human umbilical vein endothelial cells

IA:

Intra‐articular

IA2:

Insulinoma antigen 2

IBD:

Inflammatory bowel disease

IBMX:

3‐isobutyl‐1‐methylxanthine

ICG:

Indocyanine green

IDO:

Indoleamine 2,3‐dioxygenase

I

f

:

Funny current

IgA:

Immunoglobulin A

IgE:

Immunoglobulin E

IGF‐1:

Insulin‐like growth factor‐1

IGF‐1R:

Insulin‐like growth factor 1 receptor

IGFBP5:

Insulin‐like growth factor binding protein 5

IgM:

Immunoglobulin M

IL:

Interleukin

IL‐1R:

Interleukin 1 receptor

IL1RA:

Interleukin‐1 receptor antagonist protein

ILD:

Interstitial lung disease

INF‐γ:

Interferon‐ƴ

IPCs:

Insulin‐producing cells

IPF:

Idiopathic pulmonary fibrosis

iPSCs:

Induced pluripotent stem cells

IRAP:

Interleukin‐1 receptor antagonist protein

IRE:

Immunosuppressant responsive enteropathy

IRE1:

Inositol‐requiring enzyme 1

IRI:

Ischemic‐reperfusion injury

IRIS:

International Renal Interest Society

IRS‐1:

Insulin receptor substrate 1

ISCT:

International Society for Cellular Therapy

Isl‐1:

Islet‐1

ITS:

Insulin, transferrin, and selenous acid

IV:

Intravenous

IVDD:

Intervertebral disc disease

IVRT:

Isovolumic relaxation time

JNK:

Jun N‐terminal kinase

KCl:

Potassium chloride

KCS:

keratoconjunctivitis sicca

KFDA:

Korea Food Drug Administration

KLF3‐AS1:

Krüppel‐like factor 3 antisense RNA 1

Klf4:

Krüppel‐like factor 4

Ksp‐cadherin:

Kidney‐specific cadherin

L:

Lumbar

LAD:

Left anterior descending

LcHS:

Large canine hepatocyte spheroids

LDL:

Low‐density lipoprotein

LGALS9:

Galectin‐9

LIES:

Low‐intensity electrical stimulation

LIF:

Leukemia inhibitory factor

lncRNA:

Long noncoding RNA

LOCC:

Longitudinally oriented collagen conduit

LPS:

Lipopolysaccharide

LSCD:

Limbal stem cell deficiency

Lum:

Lumican

LVEF:

Left ventricular ejection fraction

LVESV:

left ventricular end systolic volume (LVESV)

M1:

Macrophage phenotype 1

Mabs:

Monoclonal antibodies

MafA:

MAF BZIP transcription factor A

MAP2:

Microtubule‐associated protein 2

MAPK:

Mitogen‐activated protein kinase

MBP:

Myelin basic protein

MCP‐1:

Monocyte chemoattractant protein‐1

MCV:

Motor conduction velocity

MDA:

Malondialdehyde

mECs:

Mouse visceral endoderm‐like cells

MEF‐2:

Myocyte enhancer factor‐2

MEFM:

Neurafilament medium chain

MEP:

Motor‐evoked potential

MF‐20:

Sarcomeric myosin

MFAT:

Microfragmented adipose tissue

mg/dL:

Milligram per deciliter

mg/kg b. wt.:

Milligram per kilogram of body weight

mg/mL:

Milligrams/milliliters

Mg:

Magnesium

MHC:

Major histocompatibility

MI:

Myocardial infarction

MIPO:

Minimally invasive plate osteosynthesis

miRNAs:

Micro ribonucleic acid

MIRO1:

Mitochondrial Rho GTPase1

MKX:

Mohawk

MLC‐2v:

Myosin regulatory light chain 2

MLR:

Mixed lymphocyte reaction

mM:

Millimolar

mmol/L:

Millimoles per liter

MMPs:

Matrix metalloproteinases

MNCs:

Mononuclear cells

MOI:

Multiplicity of infection

MPa:

Mega pascals

MPO:

Myeloperoxidase

MPSS:

Methyl prednisolone sodium succinate

MR:

Magnetic resonance Imaging

MRC1:

Mannose receptor C‐type 1

mRNAs:

Messenger ribonucleic acid

MRP:

Multidrug resistance‐associated protein

MSCs:

Mesenchymal stem cells

MSCV:

Murine stem cell virus

mShox2:

Mouse short stature homeobox 2

MST:

Multilayer sliced tendon

MUO:

Meningo‐encephalitis of unknown origin

MVI:

Mitral valve insufficiency

MVs:

Microvesicles

mW/cm

2

:

Milliwatts per square centimeter

Myf‐5:

Myogenic factor‐5

MYH9:

Myosin heavy chain 9

MyoD:

Myoblast determination protein

n:

number of animals per study or number or studies as indicated

NBBM:

Natural bovine bone mineral

NBM:

Natural bovine bone mineral

NCAM1:

Neural cell adhesion molecule 1

NEAA:

Non‐essential amino acids

NEFL:

Tau neurofilament light peptide

NES:

Nestin

NETs:

Neutrophil extracellular traps

NeuN:

Neuronal nuclear antigen

NeuroD1:

Neurogenic Differentiation 1

NF‐H:

Neurofilament H

NF‐kB:

Nuclear Factor Kappa B

NGCs:

Nerve guidance conduits

NGF:

Nerve growth factor

NGF‐b:

Nerve growth factor‐b

NGFR:

Nerve Growth Factor Receptor

Ngn3:

Neurogenin 3

nHA:

nano hydroxyapatite

nHAC:

Nano HA and collagen

NIC:

Nicotinamide

NIS:

Adenovirus‐mediated sodium iodide symporter

Nkx2.2:

NK2 Homeobox 2

NKx2‐5:

NK2 homeobox 5

NLR:

Neutrophil‐to‐lymphocyte ratio

Nm:

Nanomolar

NO:

Nitric oxide

NOD2:

Nucleotide‐binding oligomerization domain‐containing protein 2

NPH:

Neutral protamine Hagedorn

NRE:

Non‐responsive enteropathy

NSAIDs:

Non‐steroidal anti‐inflammatory drugs

NSCs:

Neural stem cells

NT‐3:

Neurotrophin 3

NT‐BNP:

N‐terminal pro b‐type natriuretic peptide

NTR:

Neurotrophin receptor

OA:

Osteoarthritis

OC:

Osteocalcin

Oct4:

Octamer‐binding transcription factor 4

Oct‐4:

Octamer‐binding transcription factor 4

OECs:

Olfactory ensheathing cells

OMEC:

Oral mucosal epithelial cell

OPCs:

Oligodendroglia precursor cells

OSM:

Oncostatin M

PAMPs:

Pathogen‐associated molecular patterns

Pax4:

Paired box 4

Pax‐7:

Paired box 7

PB:

Peripheral blood

PBS:

Phosphate buffer saline

PCBM:

Particulate cancellous bone and marrow

PCL:

Polycaprolactone

PCL‐TCP:

Polycaprolactone‐tricalcium phosphate

PCNA:

Proliferating Cell Nuclear Antigen

PCSK1:

Proprotein convertase subtilisin/kexin type 1

PD:

Periodontal disease

PDGF:

Platelet derived growth factor

PDGFRB:

Platelet‐derived growth factor receptor beta

PDL:

Periodontal ligament

PDX1

:

Pancreatic and duodenal homeobox 1

PEP:

Pancreatic endocrine progenitors

PEP:

Pre‐ejection period

PERK:

Protein kinase RNA‐like ER kinase

PGE2:

Prostaglandin E2

PI3K/Akt:

Phosphatidylinositol 3‐kinase/protein kinase B

PI3K:

Phosphatidylinositol 3‐kinase

PIS:

Pain Interference Scores

PLAC:

Poly

L

‐lactide/e‐caprolactone tube

PLB:

Phospholamban

PLE:

Protein losing enteropathy

PLGA:

Poly(lactic‐co‐glycolic acid)

PLLA:

Poly(

L

‐lactic acid)

PLR:

Platelet‐to‐lymphocyte ratio

PM:

Self‐assembling peptide nanomaterial

PMMA:

Poly(methyl methacrylate)

PMP:

Polymethylpentene

PNI:

Peripheral nerve injury

PNS:

Peripheral nervous system

Poly I:C:

Polyinosinic:polycytidylic acid

poly(PAAm‐co‐BMA:

Poly(N‐isopropylacrylamide‐co‐n‐butyl methacrylate)

POSTN:

Periostin

PP:

Polypropylene

PPARγ:

Peroxisome proliferator‐activated receptor gamma

PPRs:

Pattern recognition receptors

PRF:

Platelet‐rich fibrin

PRP:

Platelet rich plasma

PSAG:

Polysulfated glycosaminoglycan

PSS:

Pain Severity Scores

PTF:

Pre‐corneal tear film

PTGIS:

Prostaglandin I2 synthase

PTHRs:

Parathyroid hormone receptors

PU/PD:

Polyuria, polydipsia

Puppy DT:

Puppy deciduous teeth

PVAO:

Peak velocity of aortic flow

PVAS:

Pruritus visual analogue scale

PVF:

Peak vertical force

PYGM:

Glycogen phosphorylase, muscle associated

QoL:

Quality of life

RA:

Retinoic acid

RA:

Right atrium

RAA:

Renin–angiotensin–aldosterone

REX1:

Reduced expression 1

Rfx6:

Regulatory factor X6

rhBMP:

recombinant human bone morphogenetic protein

ROM:

Range of motion

ROS:

Reactive oxygen species

RTA:

Road traffic accident

RTA’s:

Road traffic accidents

RTECs:

Renal tubular epithelial cells

RT‐PCR:

Reverse transcription‐polymerase chain reaction

RUNX2:

Runt‐related transcription factor 2

RV:

Right ventricle

RVC:

Reticulated vitreous carbon

RyR:

Ryanodine receptor

SAN:

Sinoatrial node

SAP‐90:

Post‐synaptic protein

SCI:

Spinal cord injury

SCPs:

Stem cell‐based products

SCRG1:

Scrapie responsive gene 1

SCs:

Schwann cells

SCX:

Scleraxis

SDF‐1:

Stromal cell‐derived factor‐1

SDFT:

Superficial digital flexor tendon

SEP:

Somatosensory‐evoked potential

SERCA2a:

Calcium adenosine triphosphatase

SFM:

Serum free media

Sfrp2:

Secreted frizzled‐related protein 2

SHOX2:

Short Stature Homeobox 2

SII:

Systemic immune‐inflammation index

SIRS:

Systemic inflammatory respiratory syndrome

SIS:

Small intestinal submucosa

SMA:

α‐smooth muscle actin

SNPs:

Single nucleotide polymorphisms

Sox:

SRY Box Transcription factor

SOX2:

Sex‐determining region Y box 2

SPECT:

Single positron emission computed tomography

STAT3:

Signal Transducer and Activator of Transcription 3

STC1:

Stanniocalcin‐1

STT:

Schirmer tear test

SVF:

Stromal vascular fraction

SVZ:

Sub ventricular zone

Sy:

Synovial

SYP:

Synaptophysin

SYT1:

Synaptotagmin‐1

T:

Thoracic

T1D:

Type 1 diabetes

T3:

Tri‐iodothyronine

TAGLN:

Transgelin

Tagln:

Transgelin

Tbx18:

T‐box gene family

TBX3:

T‐Box Transcription Factor 3

Tbx3:

T‐Box Transcription Factor 3

TE:

Tissue engineered

TEK:

Tyrosine kinase endothelial

TEM:

Transmission electron microscopy

TERT:

Telomerase reverse transcriptase

TF:

Tissue factors

TFBC:

Tendon–fibrocartilage–bone composite

TGF‐β:

Transforming growth factor (TGF)‐β

TH1:

T helper 1 cell

THRs:

Thyroid hormone receptors

TiAl6V4:

Alpha‐beta titanium alloy

TIMP:

Tissue inhibitor matrix metalloproteinases

TLRs:

Toll‐like receptors

TNF:

Tumor necrosis factor

Tnmd:

Tenomodulin

TO‐GCN:

Time‐Ordered Gene Co‐expression Network

TPI%:

Total pressure index percentage

TPLO:

Tibial plateau leveling osteotomy

Tregs:

T regulatory cells

TrkC:

Tropomyosin receptor kinase C

TSA:

Trichostatin A

TSG:

Tumor necrosis factor‐inducible gene 6 protein

TSP2:

Thrombospondin

TSPCs:

Tendon stem/progenitor cells

TTA:

Tibial tuberosity advancement

TUBB3:

tubulin beta 3 class III

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick‐end labelling

UC:

Umbilical cord

UCB:

Umbilical cord blood

USG:

Ultrasonography

USPRS:

Ultrasound shoulder pathology rating scale

UTI:

Urinary tract infection

UTMD:

Ultrasound‐targeted microbubble destruction

VAS‐loc:

Visual Analogue Scale for locomotion

VAS‐pain:

Visual Analogue Scale for pain

VCAM‐1:

Vascular cell adhesion molecule‐1

VEGF:

Vascular endothelial growth factor

VEGFR2:

Vascular endothelial growth factor receptor 2

VI:

Vertical impulse

VLA‐4:

Very late antigen 4

vWF:

von Willebrand’s factor

WJ:

Wharton’s jelly

Wnt:

Wingless‐related integration site

α‐SMA:

Alpha‐smooth muscle actin

β1‐Ad:

β1‐adrenergic receptor

β‐TCP:

β‐tricalcium phosphate

About the Companion Website

This book is complemented by a companion website:

www.wiley.com/go/caninestemcell 

This website includes:

Videos

1Canine Mesenchymal Stem Cells Sources, Properties, and Regulations for Clinical Use

1.1 Introduction

The stem cell concept is fundamental to biology that tries to understand the development of life and the formation and function of various tissues and organs. This concept stands at the forefront of 21st‐century research and holds promise for groundbreaking advancements in biology and medicine. Stem cell research is anticipated to open new horizons, offering insights into the intricate processes of life and presenting innovative solutions in healthcare. The stem cell concept sees its inception in embryology (Gugjoo 2020). An individual, right from its initial developmental phase as a zygote to its maturity, harbors varied stem cell types. In dogs, prenatal development occurs in three phases/periods, namely, ovum period (2–17 days), embryo period (19–35 days), and fetal period (35th day to birth). In the ovum period, fertilization is followed by the development of the blastocyst that attaches to the uterus. The embryo period begins with blastocyst implantation till organogenesis is completed. Finally, characteristic features of an individual and its growth occur in the fetal period (Phemister 1974). In the early developmental stage, the potential of the primitive cell (zygote) up to the morula stage, is to develop the whole organism along with the fetal membranes (totipotent) (Fig. 1.1). As the developmental stages progress, this differentiation potential invariably decreases and the cells either become restricted in their differentiation potential or are terminally differentiated. Blastocyst that comprises inner cell mass and trophoblast has the potential to form an individual supported by fetal membranes. At this stage, specification is well established as the inner cell mass (harbors embryonic stem cells, ESCs) gives rise to an individual without its fetal membranes (pluripotent) while the trophoblast develops into fetal membranes (multipotent) (Gugjoo 2020). Contrary to this, zygote or cells in the morula give rise to the whole organism and its supporting membranes without any specification. Once the individual is born, there is a reserve cell pool in each tissue/organ that has the potential to maintain homeostasis. These adult stem cells are mostly multipotent and, in some cases, show features of pluripotency. In a particular organ/tissue, these adult stem cells get destined toward a particular lineage and the progenitor cells developed thereafter are oligopotent, which further get destined to a particular cell type (unipotent) and finally differentiate into a specialized somatic cell. Apart from the naturally available stem cells, somatic cells are also dedifferentiated into the stem cells through incorporation of the specific self‐renewal and pluripotency maintenance markers (Oct4, Sox3/4, c‐MYC, and Klf4). These adult cells induced to have pluripotency features are termed as induced pluripotent stem cells (iPSCs) (Gugjoo et al. 2020a, b, c) (Fig. 1.1).

Fig. 1.1 Sources of stem cells from initial fertilization to the adulthood of dogs. The stem cells demonstrated are totipotent (zygote/morula), pluripotent (embryonic stem cells/induced pluripotent stem cells), and multipotent (mesenchymal stem cells). The image also depicts stromalness and differentiation properties of mesenchymal stem cells.

Stem cells are being used for drug testing, understanding the development of an individual (embryology), and more recently for therapeutics. Because the focus of this book is on clinical applications of stem cells in canine practice, this chapter shall focus on the therapeutic properties of mesenchymal stem cells (MSCs) vis‐a‐vis available dog literature. ESCs or iPSCs have teratogenic potential as these cells tend to differentiate spontaneously. Additionally, ESCs have ethical issues of embryo destruction, and above all, the clear understanding of their physiological processes is still limited. Despite the challenges, there is decent literature on the utilization of ESCs or iPSCs for therapeutics in humans. However, in dogs such literature is very limited. This may be due to the above‐cited reasons; besides, the supporting infrastructure is limited and the cost of treatment with such cells remains less economical. Contrarily, adult stem cells have numerous sources, easy isolation processes, and cultivation. Besides, the teratogenic potential and ethical issues are less. Among these adult stem cell types, MSCs are seen as the most promising stem cells due to their broader sources, ease of isolation and proliferation, limited chances of teratogenicity, and ethical issues, besides their therapeutic properties (Zuk et al. 2002; Cardoso et al. 2017). As such from here on focus shall remain on MSCs.

1.2 Understanding the Stem Cell

The stem cell is an unspecialized cell of a multicellular organism that has stemness property characterized by an ability to develop its identical copy through the “self‐renewal” and a specialized destined cell through the “differentiation” (Till et al. 1964; Ab Kadir et al. 2012; Gugjoo et al. 2020d, e). Therefore, the cell that self‐renews its pool and differentiates into particular lineage as per the microenvironment/niche is a stem cell. Self‐renewal property differentiates stem cells from progenitor cells as the latter cells are unable to self‐renew but do differentiate. Self‐renewal is maintained through divisional symmetry or asymmetry. Symmetric division involves the production of identical copies in which daughter cells copy the entire genome and epigenetic changes of the parent cell. The asymmetric division produces nonidentical daughter cells through divisional asymmetry or environmental asymmetry (Wilson and Trumpp 2006; He et al. 2009). Divisional asymmetry produces one similar/identical and another nonidentical copy. This occurs when the cell fate determinants get reorganized prior to the cell division. In environmental asymmetry, microenvironment/niche‐based signals induce cell division producing identical copies. One of the identical daughter cells undergoes asymmetric cell division producing nonidentical daughter cells that function as per the demand of the local tissue (Spradling et al. 2001; Ohlstein et al. 2004). Such tissue‐specific cell production is actually the differentiation process (pluripotency/multipotency) of stem cells. The microenvironment/niche maintains cell self‐renewal and simultaneously as per the signals activates cell division for differentiation as well. With the loss of microenvironment, stem cells lose their self‐renewal properties (Li and Neaves 2006; Lilly et al. 2011).

The self‐renewal potential of the cells varies with ESCs/iPSCs able to maintain growth and proliferation for longer periods while MSCs tend to have limited self‐renewal properties. Cancer cell lines are considered to grow indefinitely due to their continuous self‐renewal ability. There is little understanding in this regard, although some of the known features of the cells are attributed toward such a property. Telomerase, an enzyme that maintains telomeres (DNA regions at chromosomal ends) activity, remains higher in stem cells including MSCs and might be responsible for self‐renewal (Greenwood and Lansdorp 2003). Nonetheless, all these stem cells express elements of various pluripotency signaling pathways such as Wnt/fzd/beta‐catenin, sonic, and Hedgehog and overexpress proteins such as homeobox 4 (Hoxb4) and caudal type homeobox 4 (Cdx4) (Watt and Hogan 2000; Kyba et al. 2002; Willert et al. 2003). It is worth noting that the self‐renewal and differentiation properties are exhibited by tumor cells as well. The possible reasons for pluripotent ESCs to self‐renew for indefinite periods may be due to their specific origin and developmental stage, tightly regulated genetics and epigenetics, and highly controlled environment. Apart from these two above‐mentioned features, numerous other specific features make stem cells including MSCs a keynote topic for research and therapeutics. The subsequent part of this chapter shall specifically focus on various aspects of MSCs.

1.2.1 MSCs Stemness or Stromalness?

Stemness of stem cells pertains to their ability to self‐renew and differentiate, while simultaneously preserving crucial differentiating genes in a state of quiescence and equilibrium. In the case of MSCs, which are a type of adult stem cell, the stemness is characterized by the capacity to undergo self‐renewal and differentiate into multiple cell lineages such as adipocytes, chondrocytes, and osteoblasts. These cells lack spontaneous differentiation as commonly seen with ESCs. Despite being recognized, the understanding of these properties is limited, and there is skepticism about the existence of true stemness properties in the cells. MSCs cell cycle is regulated in such a way as to balance self‐renewal with differentiation. Compared to pluripotent stem cells, MSCs genetic and epigenetic regulation involve use of different transcription factors (Notch, Hedgehog, and bone morphogenetic protein, BMP, pathways) and growth factors [fibroblast growth factor (FGF) and transforming growth factor‐beta (TGF‐β)] to regulate self‐renewal and differentiation. In adult tissues, the microenvironment is less tightly controlled as numerous physiological processes are undertaken and wide ranges of external stimuli arise. MSCs reside in an environment that supports their tissue repair and regeneration function. Exposure to varied stimuli eventually directs them to a state of senescence after a certain number of divisions. Stem cell‐based therapeutics occur through a multitude of processes including self‐renewal and differentiation. However, the higher the rate of their doubling, the earlier they lose these properties. In order to make maximum use of MSCs, it is important to understand their stemness and establish new techniques that can preserve multipotency for long.

MSCs stemness markers are supported by only a few genes while the molecular basis, especially the key transcription factors of their stemness, is poorly understood. Unlike ESCs, where pluripotent genes such as Oct4, Nanog, klf4, and Sox2 are well‐established, MSCs lack a single key transcription factor. To understand factors involved in stemness, highly expressed genes in undifferentiated MSCs are compared with the trilineage differentiated cells. By knocking down each gene individually, no single transcription factor is involved in the complete blockade of the differentiation process and maintaining the stemness (Kubo et al. 2009). There is progressive research that tries to understand the stemness basis of MSCs. One of the studies shows that human MSCs positive for CD271 (low‐affinity nerve growth factor receptor, LNGFR or p75 neurotrophin receptor, and p75NTR), CD90 (Thy1), and CD106 (vascular cell adhesion molecule‐1, VCAM1) retains propensity of self‐renewal and multipotency (Mabuchi et al. 2013). Two novel mechanisms, namely, scrapie responsive gene 1 (SCRG1)/bone marrow stromal cell antigen‐1 (BST1) ligand‐receptor combination and cell‐cell adhesion through N‐cadherin, are considered to establish these properties (Chosa and Ishisaki 2018).

MSCs differentiate into various lineages to the extent of pluripotency in a specific ex vivo differentiation system (extended multipotency). However, in vivo studies in numerous cases show MSCs suboptimal performance with a limited engraftment rate raising concerns over the use of the word “stem” to these cells (D'souza et al. 2015). There are well‐established biological functions that contribute to their therapeutic role and are attributed to their stromal functions rather than the multipotent differentiation (Phinney and Prockop 2007; Horwitz and Dominici 2008; Murphy et al. 2013; Gugjoo et al. 2020a, c, d). MSCs stemness and proliferation properties were initially evaluated in the 1960s and 1970s that reached maturity in the 1980s and peaked in 2000 beyond which focus on such studies was reduced. Over a period of time, the consensus over the MSCs stromalness increased with simultaneous reduction in their stemness properties for their therapeutics (Fig. 1.1). Initially, the stromalness of MSCs is realized with the study demonstrating hematopoietic supportive role through release of various growth factors, chemokines, cytokines, extracellular vesicles (EVs), or microvesicles (Matthay et al. 2017; Lykhmus et al. 2019; Spano et al. 2019; Witwer et al. 2019). Down the line, these cells are termed as medicinal signaling cells changing the paradigm but keeping the acronym (Caplan and Correa 2011). Presently, the acronym of stemness and stromalness coexist although the latter property is preferably attributed to MSCs in vivo therapeutic benefits (Mastrolia et al. 2019).

Stromalness of MSCs is achieved through their secretome harboring diverse soluble factors and EVs (Maia et al. 2017). Soluble factors of MSCs secretome contain factors such as TGF‐β1 and prostaglandin E2 (PGE2), hepatocyte growth factor (HGF), indoleamine 2,3‐dioxygenase (IDO), nitric oxide (NO), vascular endothelial growth factor (VEGF), and interleukin‐68 (IL‐68). These factors arise either constitutively or after priming with the pro‐inflammatory mediators. MSCs priming with the specific microenvironment enhances their resistance and specific expression potential (Barrachina et al. 2017; de Cássia Noronha et al. 2019; Gugjoo et al. 2023). There is good evidence supporting tissue and species‐specific variation in MSCs secretome (Carrade et al. 2012). There may or may not be comparable immunomodulation and the pathways involved may also vary. Canine BM‐MSCs and AD‐MSCs show comparable immunomodulation; however, involved pathways are different (Russell et al. 2016; Chow et al. 2017). EVs are classified as per their size, namely, exosomes (50–100 nm diameter) or microvesicles (0.1–1 μm diameter). EVs carry a diverse range of factors ranging from proteins, organelles to genetic materials (DNA and RNA) (Manzoor et al. 2023). Presently the focus is shifting toward the use of these EVs as they remain devoid of stem cell‐based limitations (immunogenicity, teratoma, and limited action in harsh environments). Besides, EVs are unaffected by the freeze‐thaw process, do not require preservatives during cryopreservation, and are economical (Abraham and Krasnodembskaya 2020).

1.2.2 MSCs Plasticity

MSCs show limited ability to trans‐differentiate in comparison to the ESCs/iPSCs. Nevertheless, their trans‐differentiation occurs across the germ layers and is demonstrated in numerous studies from varied tissue sources in humans and animals including dogs (Fig. 1.1). MSCs trans‐differentiation occurs when kept in specific differentiation media, scaffolds, or mechanical forces. MSCs are at least evaluated for their tri‐lineage differentiation in line with the International Society for Cellular and Gene Therapy (ISCT) for their therapeutic purposes (Dominici et al. 2006). However, the differentiation potential goes beyond to other lineages such as myocytes/cardiomyocytes, neural‐like cells, and germ cell‐like cells. MSCs trans‐differentiation as per the available microenvironment/niche shows that their fate is not defined but rather changeable/plastic. It is worth mentioning that true trans‐differentiation remains questionable as the cells down the lineage do not always undergo characteristic ontologic processes that are involved in the formation of a particular cell lineage (Pelagalli et al. 2018). MSCs not only are plastic in their differentiation potential but also show plasticity in immunomodulation and immunoevasion as per the available microenvironment (Wang et al. 2014; Russell et al. 2016; Wang et al. 2019). Pro‐inflammatory mediators such as interferon‐ƴ (IFN‐γ), tumor necrosis factor‐α (TNF‐α), and interleukin‐2 (IL‐2) variably activate dog MSCs immunomodulatory activities (Liu et al. 2006; Poncelet et al. 2007; Russell et al. 2016).

1.2.3 MSCs Mobilization and Homing

MSCs due to their ability to “mobilize” and “home” in the distant site make their peripheral transplantation quite a feasible option for therapeutics especially at less accessible sites. Presently studies are going on to understand the mechanisms involved in the process. MSCs are assumed to follow similar steps as those of the leukocytes. For a cell to migrate, its adhesiveness to the local milieu has to be waived off followed by its directional movement to the particular site or location. The steps involved in homing include endothelial contact of the cells by tethering and rolling, resulting in deceleration of the cells into the bloodstream. Subsequently, activation of G‐protein‐coupled receptors and simultaneous activation‐dependent arrest occurs. Finally, the cells transmigrate through the endothelium and underlying basement membrane (Butcher and Picker 1996). For resident MSCs migration and homing, mobilization occurs by downregulation of their adhesion molecules initiated by cytokines and/or chemokines such as platelet‐derived growth factor (PDGF) AB and BB, and HGF (Son et al. 2006; Ponte et al. 2007; Liu et al. 2009; Baek et al. 2011). This is followed by general principles applicable to resident and transplanted cells. In the initial step of mobilization, endothelial selectins and expression of CD44 on MSCs have an important role to play in the initial step (Sackstein et al. 2008). Activation of MSCs to mobilize occurs through G‐protein‐coupled receptors that are typically chemokine receptors. For BM‐MSCs homing, CXCR4‐stromal‐derived factor‐1 (SDF‐1) axis is critical (Moll and Ransohoff 2010; Angelone et al. 2017). Integrins play an important role in stable activation‐dependent arrest of MSCs. The integrins form dimers that bind with endothelial cell adhesion molecules. Very late antigen‐4 (VLA‐4), formed by combination of integrin α4 and β1, interacts with the VCAM‐1 and is demonstrated to be involved in MSCs homing (Rüster et al. 2006; Segers et al. 2006). Diapedesis or transmigration of MSCs through the endothelial cell layer and basement membrane is brought about by lytic enzymes such as matrix metalloproteinases (MMPs). Among various MMPs, MMP‐2 and MMP‐9 are shown to have an important role as these enzymes preferentially degrade basement membrane components (collagen and gelatin) (Nagase and Woessner 1999; Steingen et al. 2008). Finally, MSCs recruitment/homing into the inflamed and damaged tissues may be brought by adhesion molecules such as galectin‐1 and galectin‐3 (Reesink et al. 2017).

1.2.4 Anti‐inflammatory and Immunomodulatory Actions

MSCs are immune‐privileged/immunoevasive and bring in the immunomodulatory and/or anti‐inflammatory actions as dictated by the available microenvironment (Gugjoo et al. 2019a, b; Gugjoo et al. 2020a; Gugjoo and Pal 2020). Their immunoevasive nature, ability to avoid immune rejection, arises out of a lack of expression of the major histocompatibility complex (MHC)‐II and minimal expression of the MHC‐I and co‐stimulatory molecules. Immunomodulatory actions may occur either through cell–cell contact, by their secretome or constitutively. Cell–cell contact‐mediated actions arise through various MSCs receptors such as toll‐like receptors (TLRs), intracellular adhesion molecules (ICAMs), VCAM, and Fas ligand‐dependent interactions (Tomchuck et al. 2008; DelaRosa and Lombardo 2010; Akiyama et al. 2012). Transwell assays in canines show MSCs ability to secrete soluble factors that inhibit the immune response of the lymphocytes (Kang et al. 2008; Lee et al. 2011). Secretome‐based immunomodulation/inhibition seems more potent as compared to the cell–cell contact interactions (Di Nicola et al. 2002). These cells modulate CD8+ CD4+ T and dendritic cells and disrupt natural killer cells to prevent T‐cell responses (Di Nicola et al. 2002; Wang et al. 2009). MSCs interaction with the CD4 and CD8 lymphocytes is activated in the inflammatory environment that bring in the higher anti‐inflammatory actions. MSCs co‐cultured with peripheral blood‐derived mononuclear cells (PB‐MNCs) and macrophages or under pro‐inflammatory mediators (PGE2, lipopolysaccharide, interferon‐ƴ, tumor necrosis factor‐α, and IL‐2) reduce mononuclear cells (MNCs) production of inflammatory mediators such as TNF‐α, inducible nitric oxide synthase (iNOS), and interleukin (IL)‐1β (Singer and Caplan 2011; Russell et al. 2016; Chae et al. 2017; Lara et al. 2017; de Moraes et al. 2016).

1.2.5 Antimicrobial Actions

It is now a well‐accepted concept that MSCs may prevent microbial infections as has also been reported in numerous studies including the dogs (Johnson et al. 2017). Such an action occurs either indirectly (modulate local environment and/ or amplify immune cell function) or directly (eradicate microbes through antimicrobial peptides, AMPs) (Cortés‐Araya et al. 2018; Taghavi‐Farahabadi et al. 2021; Silva‐Carvalho et al. 2022). MSCs and/or secretomes may also potentiate antibiotic actions and could be useful against the biofilms (Johnson et al. 2017