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KMID : 1177720120060010035
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Journal of Alternatives to Animal Experiments
2012 Volume.6 No. 1 p.35 ~ p.40
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Comparative Study on the Formation of Murine Embryoid Body
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Shin Hyo-Sook
Seo Jong-Hun
Jeong Sang-Hee
Park Joong-Won
Son Seong-Wan
Kang Hwan-Goo
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Abstract
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The formation of embryoid body (EB) is the primary step for the differentiation of embryonic stem cells (ESCs). An EB consists of ectodermal, mesodermal and endodermal tissues, which recapitulate many aspects of cell differentiation. For this reason, EB formation has been widely utilized as a trigger of In vitro differentiation of ESCs. In this study, we established the three modified EB formation methods which were Hanging Drop (HD) method, Conical Tube (CT) method and Low Attachment (LA) Method and made a comparative study of characterization and formation efficacy of EB. There was no big difference in EB formation efficacy by inducing methods. However, more EBs expressed heart beating at HD method. Immonostaining data and EM examination results showed that EB was consist of non-differentiated ESC, variable type of differentiated cell and typical cyst. Conclusively, we suggest that EB formation by HD method is the most effective method and this protocol can be used for the developing toxicity screening of new drug.
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KEYWORD
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Mouse embryonic stem cell, Embryoid body, Hanging Drop method, Conical Tube method, Low Attachment Method
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