The Long-Term Differentiation of Embryonic Stem Cells into Cardiomyocytes: An Indirect Co-Culture Model |
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Authors: | Dong-Bo Ou Di Zeng Yan Jin Xiong-Tao Liu Ji-Wei Teng Wan-Gang Guo Hong-Tao Wang Fei-Fei Su Yong He Qiang-Sun Zheng |
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Institution: | 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.; University of Udine, Italy, |
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Abstract: | BackgroundEmbryonic 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 Millicell™ hanging cell culture inserts to control the long-term differentiation of ESCs into CMs.Methodology/Principal FindingsMouse 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 µm 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/SignificanceThese results indicate that NCMs co-culture promote ESC differentiation and has a further effect on cell growth and differentiation. We assume that the improvement of the differentiating efficiency of ESCs into CMs in the co-culture system do not result from the effect of co-culture directly on cell differentiation, but rather by signaling effects that influence the cells in proliferation and long-term function maintenance. |
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