EVALUATING THE PROLIFERATION OF CRYOPRESERVED BOVINE HAIR FOLLICLE DERIVED CELLS BETWEEN PASSAGES

Abstract

Hair follicle stem cells (HFSCs) represent a promising and readily accessible source of mesenchymal stem cells (MSCs), known for their multipotent characteristics and non-oncogenic features, which make them ideal candidates for various regenerative medicine applications. Cryopreservation is a critical technique that enables the long term storage of these cells, preserving their viability for future research and therapeutic use. This study focused on evaluating the viability, morphology, and growth characteristics of cryopreserved bovine hair follicle-derived cells (bHFCs) during in vitro expansion following the thawing process. After analysis of multiple passages, revealed significant morphological changes, including cytoplasmic and nuclear enlargement, particularly in later passages, which are indicative of cellular senescence. The study also observed an increase in apoptosis rates and a corresponding decrease in cell viability as the passages progressed. Flow cytometry analysis further indicated a G2/M phase arrest in the later passages, suggesting a disruption in the normal cell cycle progression. Additionally, Realtime qRT-PCR analysis demonstrated strong expression of key MSC markers, such as CD44, CD73, CD90, and CD105 with a significant increase in expression at later passage. These molecular markers confirmed the preservation of essential MSC characteristics in the cryopreserved bHFCs, despite the onset of senescence and other changes. The results of this study highlight the potential of cryopreserved bHFCs as a viable and valuable resource for regenerative medicine, offering a stable and multipotent cell source for future therapeutic applications. This underscores the importance of cryopreservation in maintaining the functionality of HFSCs over extended periods, ensuring their continued relevance in stem cell research and future applications.