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Acta Pharmacologica Sinica 2007 December; 28 (12): 1931-1937; doi: 10.1111/j.1745-7254.2007.00715.x |
| Original Article | [ Full text ] |
| Selective enrichment of hepatocytes from mouse embryonic stem cells with a culture system containing cholestatic serum1 |
Jun MIN2,3, Chang-zhen SHANG2, Ya-jin CHEN, Lei ZHANG, Lu LIU, Xiao-geng DENG, Mei YANG, Dong-ping CHEN, Jun CAO, Er-wei SONG, Ji-sheng CHEN Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China |
Methods: One week after the induction of E14 mouse ES cells into hepatocytes with sodium butyrate, cholestatic serum was added into the culture system at various concentrations and hepatocyte-like cells were induced to proliferate. The morphological and phenotypic markers of hepatocytes were characterized using light microscopy, immunocytochemistry, and RT-PCR, respectively. The function of glycogen storage of the differentiated cells was detected by Periodic acid-Schiff (PAS) reaction, and the ratio of hepatic differentiation was determined by counting the albumin and PAS-positive cells.
Results: In the presence of conditional selective medium containing cholestatic serum, numerous epithelial cells resembling hepatocytes were observed. The RT-PCR analysis showed that undifferentiated ES cells did not express any hepatic-specific markers; however, in the presence of sodium butyrate and conditional selective medium containing cholestatic serum, hepatic differentiation markers were detected. Immunofluorescence staining showed that those ES-derived hepatocytes were α-fetoprotein, albumin, and cytokeratin 18 positive, with the ability of storing glycogen. Further determination of the hepatic differentiation ratio showed that the application of cholestatic serum efficiently enriched ES-derived hepatocyte-like cells by inducing lineage differentiation and enhancing lineage proliferation.
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Keywords: embryonic stem cells; sodium butyrate; hepatocyte; cholestatic serum; differentia-tion |
| 1 This work was supported by research grants from the National Natural Science Founda-tion of China (No 30271277 and 30571805 to Jun MIN, No 30471799 to Xiao-geng DENG, and No 30672036 to Ya-jin CHEN). |
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