Establishing a deeper understanding of the osteogenic differentiation of monolayer cultured human pluripotent stem cells using novel and detailed analyses

Abstract Background Derivation of osteoblast-like cells from human pluripotent stem (hPSCs) is a popular topic in bone tissue engineering. Although many improvements have been achieved, the low induction efficiency because spontaneous differentiation hampers their applications. To solve this problem, detailed understanding osteogenic process hPSCs urgently needed. Methods Monolayer cultured embryonic and human-induced were differentiated commonly applied serum-containing medium for 35 days. In addition to traditional assays such as cell viability detection, reverse transcription-polymerase chain reaction, immunofluorescence, alizarin red staining, we also studies counting, telomerase activity, flow cytometry essential indicators analyse type changes each week. Results The population was quite heterogeneous throughout days induction. Then, activity cycle analyses value evaluating tumourigenicity obtained cells. Finally, dynamic map made integrate analysis these results during hPSCs, types at defined stages concluded. Conclusions Our lay foundation improve vitro by supplementing with functional compounds desired stage, then establishing stepwise system future.