心肌组织特异性表达.pdfVIP

The Long-Term Differentiation of Embryonic Stem Cells into Cardiomyocytes: An Indirect Co-Culture Model 1. 1. 2 1 1 1 1 Dong-Bo Ou , Di Zeng , Yan Jin , Xiong-Tao Liu , Ji-Wei Teng , Wan-Gang Guo , Hong-Tao Wang , 1 1 1 Fei-Fei Su , Yong He , Qiang-Sun Zheng * 1 Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi’an, China, 2 Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi’an, China Abstract Background: Embryonic Stem Cells (ESCs) can differentiate into cardiomyocytes (CMs) in vitro but the differentiation level from ESCs is low. Here we describe a simple co-culture model by commercially available MillicellTM hanging cell culture inserts to control the long-term differentiation of ESCs into CMs. Methodology/Principal Findings: Mouse ESCs were cultured in hanging drops to form embryoid bodies (EBs) and treated with 0.1 mmol/L ascorbic acid to induce the differentiation of ESCs into CMs. In the indirect co-culture system, EBs were co- cultured with epidermal keratinocytes (EKs) or neonatal CMs (NCMs) by the hanging cell culture inserts (PET membranes with 1 mm pores). The molecular expressions and functional properties of ESC-derived CMs in prolonged culture course were evaluated. During time course of ESC differentiation, the percentages of EBs with contracting areas in NCMs co-culture were significantly higher than that without co-culture or in EKs co-culture. The functional maintenance of ESC-derived CMs were more prominent in NCMs co-culture model. Conclusions/Significance: These results indicate that NCMs co-culture p