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| Stirred Bioreactor Cultivation Enhances the Efficiency of Embryoid Bodies Formation and Differentiation into Cardiomyocytes |
| HE Wen-jun,YE Ling-ling,LI Shi-chong,LIU Hong,WANG Qi-wei,WANG Hai-tao,XIE Jing,CHEN Zhao-lie |
| Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China |
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Abstract Objective: To determine the optimal condition for mouse embryonic stem cells (mESC) culture with stirred bioreactor, and to develop a method for mass production of embryoid bodies (EB). Methods: The different initial cell concentrations of mESC and the initial stirring speed of bioreactor were investigated to determine the optimal condition for EB formation. Induced by ascorbic acid, the differentiation of EBs formed in stirred bioreactor into cardiomyocytes was compared with EBs formed in Petri dish. Immunofluorescence staining and RTPCR were used to identify the cardiomyocytes derived from mESC. Results: The formation of a large number of uniform relatively EBs was achieved in stirred bioreactor when mESC were seeded initially with 1×105~3×105 cells/ml and stirring speed was set to 15~30r/min. Most of cells in the EBs formed in bioreactor were viable. EBs produced in bioreactor differentiated into cardiomyocytes more efficiently compared with EBs from Petri dish. The cardiac specific genes were expressed in ESCderived cardiomyocytes. Conclusions: Stirred bioreactor culture could enhance the efficiency of EB formation and differentiation into cardiomyocytes, which may be a more ideal culture system for EB formation.
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Received: 14 July 2009
Published: 07 December 2009
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