Ratan K Choudhary

Title : Harnessing canine stromal vascular fractions: Unraveling immunomodulation in co-culture with PBMCs for enhanced tissue repair and regeneration

Abstract:

In recent years, mesenchymal stem cell (MSC) therapy has been investigated in dogs, but inconsistent outcomes have been observed. Stromal Vascular Fractions (SVFs) present a heterogeneous cell population, including MSCs, progenitor cells, endothelial cells, pericytes, and immune cells, with potential advantages for tissue repair and regeneration over isolated MSCs. This study aimed to analyze SVFs' growth dynamics and assess their immunomodulatory efficacy in co-culture with PBMCs and allogeneic SVFs. SVFs were isolated from healthy female dogs (n = 3) undergoing cesarean section, with owner consent, while PBMCs were collected from tumor-free and lymphoma dogs (n = 5 each). Quantitative evaluation of SVFs cell proliferation and gene expression analysis (OCT4, CD44, CD90, and CD105) was performed in a 3-day culture. Next, SFVs were co-cultured with PBMCs (of healthy and lymphoma dogs) at different ratios (1:1, 1:0.5, and 1:0.1; N/N) for three days, and the resulting co-cultures were assessed for cell proliferation. Results showed canine SVFs expressed cell surface markers CD105+/CD90+/CD44+ and pluripotency transcription factor OCT4. Co-culturing SVFs and PBMCs resulted in increased population doubling time (PDT) in both the PBMCs and decreased Ki67 expression in the lymphoma group. Cell proliferation in co-culture decreased in a ratio-dependent manner. Immunomodulatory properties of canine SFVs were evident by the down-regulation (44x; p-value = 0.0003) of the inflammatory cytokine TNFA and up-regulation (88x; p-value = 0.008) of the anti-inflammatory cytokine PTGS1 in PBMCs in a mixed culture of SVFs (1:1 ratio). In conclusion, canine SVFs derived from periovarian fatty tissue exhibited a heterogeneous cell population, including mesenchymal stem cells and immune cells. Co-culture with PBMCs demonstrated immunomodulatory effects, complementing their potential for therapeutic applications. The expression of key stem cell markers further supports their therapeutic promise.

Audience Take Away:

• The audience will know about stem cell research and its utility in canine regenerative medicine.
• The practical utility of this research lies in the potential application of Stromal Vascular Fractions (SVFs) in mesenchymal stem cell (MSC) therapy for dogs. The study shows SVFs have advantages over isolated MSCs for tissue repair and regeneration.
• A methodology for SVF isolation from canine adipose tissue

Biography:

Dr. Ratan K. Choudhary is an Assistant Professor at the College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Punjab, India. With BVSc from TNVASU, Chennai, and a master's degree in Animal Genetics and Breeding from IVRI, Bareilly, he earned a doctoral degree in Animal Science from the University of Maryland, USA. Dr. Choudhary's research focused on bovine mammary stem cells, enhancing lactation persistency in cattle. Having worked at the USDA facility for over six years, he gained valuable experience in stem cell research during two post-doctoral positions at the University of Kentucky and the University of Vermont, USA. His long-term research goal is to develop stem cell therapies for dairy animals, including mastitis treatment. He has been recognized with numerous awards and fellowships for his research work and has published three books and more than 50 papers in international peer-reviewed journals. Dr. Choudhary is an active member of various international scientific societies and organizations, contributing significantly to the field of stem cells in veterinary sciences.