Fig. 3
Mass production of AM-MSCs. a The isolation procedure, and the number of established AM-MSCs per cm2 of the amniotic membrane. Y-axis values represent the logarithm of total cell numbers. b Growth rate of AM-MSCs based on passages (Biological replication, n = 3). c mRNA levels of pluripotency-related genes OCT4, SOX2, and NANOG according to the passage number. GAPDH was used as an internal control for RT-qPCR (Technical replication, n = 3). d Expression percentage of MSC markers (CD90, CD105) and hematopoietic markers (CD34) in early (p.2) and late (p.10) passage of AM-MSCs. e mRNA expression analysis of osteogenesis-related (ALP, OCN) and adipogenesis-related (PPARγ, FABP4) genes on day 14 after the induction of differentiation (Biological replication, n = 3). GAPDH was used as an internal control. P-values < 0.05 were considered significant. *** P < 0.001. undif, undifferentiated cells; dif, differentiated cells. f Estimation of total cell numbers during the passage. More than 2 × 1013 AM-MSCs could be harvested after ten passages. If maximally, the 2 × 108 cells are used to transplant into a patient, it can be used in more than 10,000 people. Y-axis values represent the logarithm of total cell numbers