Expression dynamics of mesenchymal stem cell markers in canine adipose-derived stromal vascular fraction during culture

Title : Expression dynamics of mesenchymal stem cell markers in canine adipose-derived stromal vascular fraction during culture

Abstract:

The stromal vascular fraction (SVF) is a heterogeneous cell population derived from adipose tissue that has gained attention for its potential in regenerative medicine. This study investigated the expression dynamics of mesenchymal stem cell (MSCs) markers, CD90, CD105, and CD44 during in vitro expansion of canine SVF. The SVF was enzymatically isolated from the periovarian fat of healthy dogs and cultured from passage 0 (P0) to passage 3 (P3). Immunocytochemical localization of CD90 and CD105 markers was performed. Quantitative analysis of the percentage of marker positive cells revealed a 3% decrease in CD90-positive and a 10.5% decrease in CD105-positive cells from P0 to P3. In flow cytometry, the percentage of CD44+ and CD90+ cells in the P1 passage was 10.9% and 76.9%, respectively, with 20.7% of double positive CD45- cells. In the P3 passage, CD44+ and CD90+ cells were 4.1 % and 87.7 %, respectively with 1.5 % of double positive CD45- cells. The percentage of double marker positive cells (representing enriched MSCs fraction) was reduced drastically by 83.7% within two passages that is from P1 to P3 from 20.7 % to 1.5%. These findings indicate a gradual phenotypic shift during serial passaging, which could impact the therapeutic potential of SVF- derived MSCs. A reduction in hematopoietic lineage negative (CD45-) and stem cell marker (CD44+, CD90+, and CD105+) positive cells may reflect a loss of regenerative capability over time. Notably, the CD90 was abundantly expressed in canine SVF in all fat samples. This study underscores the importance of utilizing freshly isolated or early-passage SVF for cell-based therapies to maximize stem cell functionality and ensure therapeutic efficacy. Our findings provide valuable insights into the phenotypic stability of canine adipose tissue derived SVF.

Biography:

Hitesh Rana is a PhD student working in the Animal Stem Cell Lab at the College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Punjab, India. His research focuses on the isolation and characterization of stromal vascular fractions (SVFs) derived from canine adipose tissue. He investigates their immunomodulatory properties through in vitro and in vivo assays to assess their potential therapeutic applications.
His work involves evaluating the regenerative capabilities of SVFs, particularly in wound healing and lymphoma treatment. This includes studying their safety profile, efficacy, and mechanisms of action in modulating the immune response. By employing advanced techniques such as immunocytochemistry, quantitative PCR, and flow cytometry, he analyze cellular markers and gene expression patterns that contribute to tissue repair and immune regulation. Beyond his laboratory work, he is passionate about translational research and the development of innovative stem cell-based therapies for veterinary and biomedical applications. He aims to bridge the gap between fundamental research and clinical implementation, contributing to the advancement of regenerative medicine.