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Section: Animal Cell Technology

Enhanced cardiac differentiation of mouse embryonic stem cells by use of the slow-turning, lateral vessel (STLV) bioreactor

Sasitorn Rungarunlert Nuttha Klincumhom Istvan Bock Csilla Nemes Mongkol Techakumphu Melinda K. Pirity Andras Dinnyes

S. Rungarunlert N. Klincumhom I. Bock Cs. Nemes MK. Pirity A. Dinnyes BioTalentum Ltd., Aulich Lajos u. 26. H-2100, Godollo, Hungary

S. Rungarunlert N. Klincumhom M. Techakumphu Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330 Thailand

I. Bock A. Dinnyes Molecular Animal Biotechnology Laboratory, Szent Istvan University, H-2100 Gdll, Hungary Corresponding author: andras.dinnyes@biotalentum.hu; Phone: +36/20/510-9632, Fax: +36/28/526-151

Emails: Sasitorn Rungarunlert nut_vs@yahoo.com Nuttha Klincumhom nuttha.klincumhom@biotalentum.hu Istvan Bock istvan.bock@biotalentum.hu Csilla Nemes csilla.nemes@biotalentum.hu Mongkol Techakumphu Mongkol.T@chula.ac.th Melinda K. Pirity melinda.pirity@biotalentum.hu

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Abstract Embryoid body (EB) formation is a common intermediate during in vitro differentiation of pluripotent stem cells into specialized cell types. We have optimized the slow-turning, lateral vessel (STLV) for large scale and homogenous EB production from mouse embryonic stem cells. The effects of inoculating different cell numbers, time of EB adherence to gelatin-coated dishes, and rotation speed for optimal EB formation and cardiac differentiation were investigated. Using 3x105 cells/ml, 10 rpm rotary speed and plating of EBs onto gelatin-coated surfaces three days after culture, were the best parameters for optimal size and EB quality on consequent cardiac differentiation. These optimized parameters enrich cardiac differentiation in ES cells when using the STLV method.

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