Isolation, Cultivation, Characterization and Expansion of Human Adipose-Derived Mesenchymal Stem Cell for Use in Regenerative Medicine

Abstract

Introduction: Stromal cells having preadipocyte characteristics can be isolated from adipose tissue, propagated in vitro and induced to differentiate in vitro toward the osteogenic, adipogenic, myogenic and chondrogenic lineages when treated with established lineage-specific factors. In this research we isolated stromal cells from human adipose tissue and cultured and expanded and examined their stemness by determining their surface CD markers and their ability to differentiate into adipocyte lineage.
Material and methods: For isolating ASCs, raw lipoaspirates were washed with sterile phosphate-buffered saline (PBS) containing 5% penicillin/streptomycin. To digest the adipose tissue, aspirates were treated with 0.075% collagenase for 1 h. To differentiate the cell to adipocyte, confluent cells were exposed to adipogenic medium containing α-MEM, FBS, dexamethasone, indomethacin, IBMX, L-glutamin, and penicillin/streptomycin.
Results: Adhered cells were cultured for 2-3 weeks with replacing the media every 3-4 days and the ADSC were isolated, cultured and expanded. To examine the differentiation potential of the isolated cell, they were differentiated in specific medium and lipid droplets were appeared within the cells within 2-3 weeks. To confirm the lipid droplet, Oil red O lipid staining was used.
Conclusion: In conclusion, it could be taken to serious consideration that, besides, other sourses of mesenchymal stem cell, adipose derived are one the best and promising source being easily accessible and available by noninvasive method, as well as potential of being used in autologous cell transplantation in wide variety of disorders from nerve to cardiac injury from one side and musculoskeletal problem from other side.