Jichu yixue yu linchuang (Aug 2021)
Establishment of a serum-free culture method for human amnion mesenchymal stem cells
Abstract
Objective To establish a method of serum free culture for human amnion mesenthymal stem cells. Methods The hAMSCs were isolated by trypsin-collagenase two step digestion method, then cultured in serum free medium. Then immunofluorescence and WST assay were used to detect cell phenotype and cells growth. Cell migration of hAMSCs cultured in serum free medium was examined by scratch wound closure assay. The expression of growth factor was detected by real time PCR assay. Results The hAMSCs cultured in serum free medium exhibited spindle-shape characteristics, were positive for CD73, CD90, but negative for CD45 and CD34. The cell proliferation showed a U-shaped curve and the average doubling time of was 72 hours.The scratch wound closure assay demonstrated a statistically significant increase of migration as compared with those cultured in FBS.The expression of VEGF,KGF,PDGF,IGF-1 was up-regulated compared with those cultured in FBS. Conclusions A serum free culture method of hAMSCs is established, which provides an experimental evidence supporting potential application.
