Chen Qijie, Kon Junko, Ooe Hidekazu, Sasaki Kazunori, Mitaka Toshihiro
Department of Pathophysiology, Cancer Research Institute, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-Ku, Sapporo 060-8556, Japan.
Nat Protoc. 2007;2(5):1197-205. doi: 10.1038/nprot.2007.118.
This protocol details a method of obtaining selectively proliferated hepatocyte progenitor cells using hyaluronic acid (HA)-coated dishes and serum-free medium. A small hepatocyte (SH) is a hepatocyte progenitor cell of adult livers and has many hepatic functions. When the rat SH begins to proliferate, CD44 is specifically expressed. To define the purification of SH, CD44 and cytokeratin 8 are used as marker proteins. The growth of SHs is faster on HA-coated dishes than on other extracellular matrix-coated ones. The use of both DMEM/F12 medium and HA-coated dishes allows the selective proliferation of SHs in culture. The purification of SHs is approximately 85% at day 10.
本方案详细介绍了一种使用透明质酸（HA）包被培养皿和无血清培养基获得选择性增殖的肝祖细胞的方法。小肝细胞（SH）是成年肝脏的肝祖细胞，具有多种肝功能。当大鼠SH开始增殖时，CD44会特异性表达。为了定义SH的纯化，CD44和细胞角蛋白8被用作标记蛋白。与其他细胞外基质包被的培养皿相比，SH在HA包被的培养皿上生长更快。使用DMEM/F12培养基和HA包被的培养皿可使SH在培养中选择性增殖。在第10天时，SH的纯化率约为85%。
