Shiojiri Nobuyoshi, Nitou Miho
Department of Biology, Faculty of Science, Shizuoka University, Shizuoka, Japan.
Methods Mol Biol. 2012;826:3-10. doi: 10.1007/978-1-61779-468-1_1.
To investigate cell-cell interactions during mammalian liver development, it is essential to separate hepatoblasts (fetal liver progenitor cells) from nonparenchymal cells, including stellate cells, endothelial cells, and hemopoietic cells. Various factors, which may be produced by nonparenchymal cells, could be assayed for their effects on the growth and maturation of separated hepatoblasts. The protocol using immunomagnetic beads coated with anti-mouse E-cadherin antibody is described for efficient isolation of hepatoblasts from cell suspensions of fetal mouse livers. The purity and recovery rate are larger than 95% and approximately 30%, respectively. The protocol may be useful for various studies focusing on the fetal liver progenitor cells.
为了研究哺乳动物肝脏发育过程中的细胞间相互作用,将成肝细胞(胎儿肝脏祖细胞)与非实质细胞(包括星状细胞、内皮细胞和造血细胞)分离至关重要。可以检测各种可能由非实质细胞产生的因子对分离出的成肝细胞生长和成熟的影响。本文描述了一种使用包被有抗小鼠E-钙黏蛋白抗体的免疫磁珠从胎鼠肝脏细胞悬液中高效分离成肝细胞的方法。纯度和回收率分别大于95%和约30%。该方法可能对各种聚焦于胎儿肝脏祖细胞的研究有用。