Culture And Osteogenic Induction Of Human Amnion Mesenchymal Stem Cells On Microcarriers

Human amnion mesenchymal stem cells (hAMCs) are able to differentiate into multiple cell lineages and display immunomodulatory properties. They are easily procured, no risks for donors, and no ethical concerns. Due to these advantages, hAMCs have great potential in clinical applications and scientific research.This research investigated the effects of cytodex3 microcarrier culture and two-dimensional (2D) static culture on expansion, metabolisms of subcultured hAMCs and self-renewal and differentiation potential of cells harvested from logarithmic phase. It was also investigated that effects of dynamic osteogenic induction culture and 2D static osteogenic induction culture on cell proliferation, metabolisms and osteogenic differentiation as well as the effects of fluid stimulation and collagen I on hAMCs proliferation and osteogenic.The results show that the attachment rate of microcarrier culture system increased to (86.3±2.5)% with intermittent stirring,4% FBS, half dose volume inoculation method, and cell number in microcarrier culture system with higher glucose utilization was about 26 times than that in 2D culture. Colony formation rate (P<0.05), calcium matrix and intracellular lipid droplets (P<0.05) of hAMCs harvested from microcarrier culture system were higher than those from 2D culture. After 14d osteogenic induction, cell number in microcarrier dynamic osteogenic induction culture system was about 26 times than that in 2D culture, and yield of lactic acid were lower than corresponding 2D static osteogenic induction culture system, alkaline phosphatase activity and extracellular matrix calcium content were higher than that in 2D static osteogenic induction culture system (P<0.05). The research of 14d osteogenic induction in 2D culture indicated that fluid stimulation and collagen I had little effect on cellular morphology, but fluid stimulation significantly inhibited the proliferation of hAMCs (P<0.05), coating collagen I had no significant role in promoting cell proliferation (P> 0.05). Both fluid stimulation and collagen I could promote alkaline phosphatase activity, calcium content, and the combined application of fluid stimulation and collagen I coating had stronger promotion on hAMCs osteogenic induction.