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The long-term differentiation of embryonic stem cells into cardiomyocytes: an indirect co-culture model.


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 Millicell™ 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 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/significance

These 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.

SUBMITTER: Ou DB 

PROVIDER: S-EPMC3557249 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Publications

The long-term differentiation of embryonic stem cells into cardiomyocytes: an indirect co-culture model.

Ou Dong-Bo DB   Zeng Di D   Jin Yan Y   Liu Xiong-Tao XT   Teng Ji-Wei JW   Guo Wan-Gang WG   Wang Hong-Tao HT   Su Fei-Fei FF   He Yong Y   Zheng Qiang-Sun QS  

PloS one 20130128 1


<h4>Background</h4>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 Millicell™ hanging cell culture inserts to control the long-term differentiation of ESCs into CMs.<h4>Methodology/principal findings</h4>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  ...[more]

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