Characterization and differentiation potential of mesenchymal stem cells isolated from multiple canine adipose tissue sources

Usman Rashid,Jörg R Aschenbach,Muhammad Yaqoob,Muhammad Moaeen-Ud-Din,Gerhard Sponder,Arfan Yousaf,Shahid Waseem,Sandra K Becker,Evelyn Saba,Mansur Abdullah Sandhu

doi:10.1186/s12917-021-03100-8

Usman Rashid, Jörg R Aschenbach + Show 8 more

Open Access

https://doi.org/10.1186/s12917-021-03100-8

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Abstract

BackgroundMesenchymal stem cells (MSCs) are undifferentiated cells that can give rise to a mesoderm lineage. Adipose-derived MSCs are an easy and accessible source for MSCs isolation, although each source of MSC has its own advantages and disadvantages. Our study identifies a promising source for the isolation and differentiation of canines MSCs. For this purpose, adipose tissue from inguinal subcutaneous (SC), perirenal (PR), omental (OM), and infrapatellar fat pad (IPFP) was isolated and processed for MSCs isolation. In the third passage, MSCs proliferation/metabolism, surface markers expression, in vitro differentiation potential and quantitative reverse transcription PCR (CD73, CD90, CD105, PPARγ, FabP4, FAS, SP7, Osteopontin, and Osteocalcin) were evaluated.ResultsOur results showed that MSCs derived from IPFP have a higher proliferation rate, while OM-derived MSCs have higher cell metabolism. In addition, MSCs from all adipose tissue sources showed positive expression of CD73 (NT5E), CD90 (THY1), CD105 (ENDOGLIN), and very low expression of CD45. The isolated canine MSCs were successfully differentiated into adipogenic and osteogenic lineages. The oil-red-O quantification and adipogenic gene expression (FAS, FabP4, and PPARγ) were higher in OM-derived cells, followed by IPFP-MSCs. Similarly, in osteogenic differentiation, alkaline phosphatase activity and osteogenic gene (SP7 and Osteocalcin) expression were higher in OM-derived MSCs, while osteopontin expression was higher in PR-derived MSCs.ConclusionIn summary, among all four adipose tissue sources, OM-derived MSCs have better differentiation potential toward adipo- and osteogenic lineages, followed by IPFP-MSCs. Interestingly, among all adipose tissue sources, MSCs derived from IPFP have the maximum proliferation potential. The characterization and differentiation potential of canine MSCs isolated from four different adipose tissue sources are useful to assess their potential for application in regenerative medicine.

Highlights

Mesenchymal stem cells (MSCs) are undifferentiated cells that can give rise to a mesoderm lineage
Infrapatellar fat pad Mesenchymal stem cells doubling time The cell doubling time of MSCs isolated from different sites was evaluated at P3 (Fig. 3A)
Our results showed that OM, SC, and PR derived MSCs have a significantly (P = 0.004) higher cellular metabolism compared to infrapatel‐ lar fat pad (IPFP)-MSCs

Summary

Introduction

Mesenchymal stem cells (MSCs) are undifferentiated cells that can give rise to a mesoderm lineage. Regenerative medicine has gained significant attention in human and veterinary medicine because of its potential role in healing or replacing apoptotic cells. MSCs can be isolated from different parts of the body, including adipose tissue [2] bone marrow, umbilical cord [3], and synovial fluid [4]. International Society for Cell Therapy recommended three basic characteristics of the regenerative potential of MSCs, including plastic adhesion, the expression of clusters of differentiation (CD) including CD73, CD90, CD105, and multi-lineage differentiation [5]. Adipose-derived mesenchymal stem cells (AD-MSCs) have attracted attention because they can be collected by minimal invasive surgery. Contrasted with bone marrow-derived MSCs (BMSCs); AD-MSCs produce more cells, have a higher proliferation rate, and maintains their phenotype during higher passages [6]. Past literature on canine species has proven the use of AD-MSCs for the treatment of osteoarthritis of the elbow [8], coxofemoral [9] and humeroradial joint [10]

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