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

Abstract

Background Mesenchymal 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. Results Our 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. Conclusion In 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.

A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources

“…Therefore, in the present study, we compared MSCs isolated from three different canine tissues (i.e., bone marrow, adipose tissue, and neonatal amniotic membrane), which have certain advantages/disadvantages in terms of isolation and yields and their biological activities eligible for the regeneration of specific tissues. Our results are consistent with other works [ 22 , 23 , 24 ] and confirm the ability to isolate MSCs from various canine adults as well as perinatal tissues.…”

Current Status and Future Prospects of Stem Cell Therapy in Animal Health

“…The results indicate that the cells from each tissue possess a unique set of features besides the basic features of stem cells and these features should be kept in consideration while choosing an appropriate source MSCs for stem cell based therapeutic purposes. In domestic and companion animals, MSCs have been isolated from their adult individuals as well as from their respective fetal annexes including bone marrow (Sasaki et al 2018;Zhang et al 2018;Arévalo-Turrubiarte et al 2019), fat tissue (Sasaki et al 2018;Arévalo-Turrubiarte et al 2019;Rashid et al 2021b), synovial fluid (Bearden et al 2017;Arévalo-Turrubiarte et al 2019), patellar fat (Rashid et al 2021a), umbilical cord (Zhang et al 2018;Denys et al 2020), Wharton's Jelly (Sarfraz et al 2021), cord blood (Kang et al 2012), periosteum and muscle (Kisiel et al 2012;Arévalo-Turrubiarte et al 2019), peripheral blood (Sato et al 2016;Longhini et al 2019), periodontal ligament and gingiva (Mensing et al 2011), placenta (Carrade et al 2011), amniotic fluid (Sarfraz et al 2021) and endometrium (Rink et al 2017). In lab animals, MSCs have also been isolated from soft tissues including the brain, liver, pancreas, kidney, spleen, thymus, lung and muscle (Meirelles et al 2006).…”

“…They differ in terms of growth kinetics, plasticity and secretory activity (Arévalo-Turrubiarte et al 2019;Fideles et al 2019;Villatoro et al 2019). Even within same type of tissue, for example adipose tissue, the anatomical location for cell isolation matters (Yaneselli et al 2018;Rashid et al 2021b). The established fact is that the phenotypic and genotypic characteristics of the stem cells are greatly influenced by the type and location of the tissue, therefore, it is of utmost importance to consider these properties while choosing the cells for regenerative and therapeutic purposes.…”

Stromal Vascular Fraction From Canine Adipose Tissue Comprises Mesenchymal Stromal Cell Subpopulations Characterized by Time-dependent Binding Affinity to Culture Plastic

“…In addition, canine MSC have been extensively studied due to the interest this species holds as preclinical/clinical models for cell therapy in humans (Hoffman & Dow, 2016). Different sources of MSC are currently used in dogs, but adipose tissue (AT) is one of the most studied since it is abundant, easy to collect using minimally invasive procedures, and rich in MSC that are relatively easy to isolate, propagate and exhibit high proliferative potential in vitro (Rashid et al, 2021;Zhan et al, 2019).…”