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首页> 外文期刊>Chinese journal of digestive diseases >Differentiation of mouse embryonic stem cells into hepatocytes in vitro and in vivo
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Differentiation of mouse embryonic stem cells into hepatocytes in vitro and in vivo

机译:小鼠胚胎干细胞体外和体内分化为肝细胞

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OBJECTIVE: Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages. Cell-to-cell contact is important for cell differentiation. Mouse ES cells were cocultured with mouse fetal liver cells and the green fluorescent protein (GFP) positive ES cells were transplanted into rats liver through the portal vein in order to investigate their potential to differentiate into hepatocytes.METHODS: Mouse ES cells were cocultured with the mouse fetal liver cell line, BNL.CL_2. They did not make direct contact; instead the culture media was exchanged freely. After coculture for 48 h, albumin, transthyretin, glucose 6 phosphates, hepatic nuclear factor 4 and SEK1 mRNA were assayed by RT-PCR, and alpha-fetoprotein by immunohistochemistry. The morphology was investigated by microscopy. After transplantion of the GFP-positive ES cells, the whole liver was removed from a rat every four days. The liver slices were examined under a fluorescent microscopeto detect the GFP-positive cells. Albumin was detected on the same slices by immunohistochemistry.
机译:目的:胚胎干(ES)细胞具有分化为多种细胞谱系的多能能力。细胞间接触对于细胞分化很重要。将小鼠胚胎干细胞与小鼠胎儿肝细胞共培养,并将绿色荧光蛋白(GFP)阳性胚胎干细胞通过门静脉移植到大鼠肝脏中,以研究其分化为肝细胞的潜力。小鼠胎儿肝细胞系BNL.CL_2。他们没有直接联系;相反,文化媒体可以自由交换。共培养48小时后,通过RT-PCR检测白蛋白,运甲状腺素蛋白,磷酸6葡萄糖,肝核因子4和SEK1 mRNA,并通过免疫组织化学检测α-甲胎蛋白。通过显微镜检查形态。移植GFP阳性ES细胞后,每四天从大鼠中取出整个肝脏。在荧光显微镜下检查肝切片以检测GFP阳性细胞。通过免疫组织化学在相同的切片上检测到白蛋白。

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