Abstract
BackgroundAdipose tissue-derived mesenchymal stem cells (ASCs) offer a promising cell source for therapeutic applications in musculoskeletal disorders. The appropriate selection of ASCs from various fat depots for cell-based therapy is challenging. The present study aims to compare stemness and multipotency of ASCs derived from retroperitoneal (RP), subcutaneous (SC), and lipoma (LP) fat to assess their usefulness for clinical application.MethodsEquine ASCs from the three fat tissue sources were isolated and characterized. The cell viability, proliferation, and self-renewal were evaluated using MTT, sulforhodamine B, and colony forming unit (CFU) assays. Stem cell relative marker CD44, CD90, and CD105 and tumor marker CA9 and osteopontin (OPN) expression were quantified using RT-qPCR. Multipotency of ASCs for adipogenic, osteogenic, and chondrogenic differentiation was examined by quantifying Oil Red O and Alizarin Red S staining, alkaline phosphatase activity (ALP), and expression of differentiation relative markers. All data were statistically analyzed using ANOVA.ResultsRP fat-derived ASCs showed a higher cell proliferation rate compared to SC and LP derived cells. In contrast, ASCs from lipoma displayed a lower proliferation rate and impaired CFU capacities. The expression of CD44, CD90, and CD105 was upregulated in RP and SC derived cells but not in LP cells. RP fat-derived cells displayed a higher adipogenic potential compared to SC and LP cells. Although ASCs from all fat sources showed enhanced ALP activity following osteogenic differentiation, SC fat-derived cells revealed upregulated ALP and bone morphogenetic protein-2 expression together with a higher calcium deposition. We found an enhanced chondrogenic potency of RP and SC fat-derived cells as shown by Alcian blue staining and upregulation of aggrecan (Aggre), cartilage oligomeric matrix protein precursor (COMP), and collagen 2a1 (Col2a1) expression compared to LP. The expression of OPN and CA9 was exclusively upregulated in the ASCs of LP.ConclusionsThe results provide evidence of variation in ASC performance not only between normal fat depots but also compared to LP cells which suggest a different molecular regulation controlling the cell fate. These data provided are useful when considering a source for cell replacement therapy in equine veterinary medicine.
Highlights
Adipose tissue-derived MSCs (ASCs) are a promising cell source in regenerative medicine in terms of their therapeutic applications for musculoskeletal disorders involving bone, cartilage, and tendon lesions [1]
While the subcutaneous fat samples (n = 8) were obtained from horses euthanized due to reasons not related to this study, samples from retroperitoneal fat (n = 8) and mesenteric lipomas (LP; n = 8) were obtained from horses undergoing abdominal surgery at the Clinic for Horses, Department of Surgery, at the Faculty of Veterinary Medicine, Justus-LiebigUniversity of Giessen
Adipose tissue-derived mesenchymal stem cells (ASCs) showed a negative immunoreaction against CD45 and CD34 harvested from RP, SC, and LP fat (Fig. 1r–w)
Summary
Adipose tissue-derived MSCs (ASCs) are a promising cell source in regenerative medicine in terms of their therapeutic applications for musculoskeletal disorders involving bone, cartilage, and tendon lesions [1]. In contrast to bone marrow, subcutaneous adipose tissue is easier to harvest and the sampling technique is associated with fewer risks [5]. Another detrimental factor is stemness, meaning the long-term self-renewal and multipotential differentiation of MSCs [6]; it has been shown that bone marrow MSCs exhibited signs of cell senescence at passage 7; ASCs can be cultivated up to passage 8 without any signs of senescence [7]. The present study aims to compare stemness and multipotency of ASCs derived from retroperitoneal (RP), subcutaneous (SC), and lipoma (LP) fat to assess their usefulness for clinical application
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
