Differentiation of embryonic stem cells into hepatocytes induced by a combination of cytokines and Trichostatin A

Zhu Xiaobo; ZHANG Dongmei; ZHANG Qian; ZHANG Jie; ZHOU Mingming

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Differentiation of embryonic stem cells into hepatocytes induced by a combination of cytokines and Trichostatin A

Zhu Xiaobo 1,ZHANG Dongmei 2,ZHANG Qian 2,ZHANG Jie 3,ZHOU Mingming 2 * #

1.Rugao Renmin Hospital,department of paediatrics , Nantong 226000

2.Medicine school, Nantong University, Nantong 226019

3.Life Science shool, Nantong University, Nantong 226007

*Correspondence author

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Funding: 如皋市科技计划SRG（No.12)300）, 南通大学博士启动基金（No.03080519）, 南通市科技计划项目（No.S2008049，BK2011015）, 江苏省博士后科研资助计划（No.1201023B）, 江苏省高校自然科学研究面上项目（No.12KJB310009）, 南通大学校级课题（No.2012002）

Opened online: 6 November 2013

Accepted by: none

Citation: Zhu Xiaobo,ZHANG Dongmei,ZHANG Qian.Differentiation of embryonic stem cells into hepatocytes induced by a combination of cytokines and Trichostatin A[OL]. [ 6 November 2013] http://en.paper.edu.cn/en_releasepaper/content/4566643

In this paper，we presented a novel 3-step procedure to efficiently direct the differentiation of mouse ESCs into hepatocytes induced by combination of cytokines and sodium butyrate. Mouse ESCs were first differentiated into definitive endoderm cells by 3 days of Activin A treatment; next, presence of acid fibroblast growth factor(aFGF)and Trichostatin A (TSA)in the culture medium for 5 days, definitive endoderm cells efficiently differentiated to hepatocytes. After 10 days of further in vitro maturation, the morphological and phenotypic markers of hepatocytes were characterized using light microscopy, immunofluorescence and RT-PCR, respectively. Furthermore, these cells were tested about the functions associated with mature hepatocytes including glycogen storage, indocyanine green uptake and release,and the ratio of hepatic differentiation was determined by counting the albumin-positive cells，which showed that the ratio of hepatic differentiation was 57.38%． The ES cell differentiating method provides a new resource for hepatocytes tansplatation．

Keywords:embryonic stem( ES) cells; hepatocyte; induced differentiation

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