Mitaka T, Kojima T, Mizuguchi T, Mochizuki Y
Department of Pathology, Sapporo Medical University School of Medicine, Japan.
Biochem Biophys Res Commun. 1995 Sep 14;214(2):310-7. doi: 10.1006/bbrc.1995.2289.
Small hepatocytes existed in the supernatant following low-speed centrifugation of the cell suspension after collagenase liver perfusion. The cells proliferated for more than 2 months and formed colonies in the Dulbecco's modified Eagle's medium supplemented with 10 mM nicotinamide, 10% fetal bovine serum, 1 mM ascorbic 2-phosphate, and 10 ng/ml epidermal growth factor. One small cell finally proliferated to several hundred cells. In addition, some cells in the colonies were shown to differentiate into mature hepatocytes that had a large cytoplasm and sometimes two nuclei. The secretion of albumin in the medium by the hepatocytes increased with time in culture, and the cells possessed connexin 32 in their cell membrane and many peroxisomes. Thus, the small hepatocytes may be "committed progenitor cells" which can further differentiate into mature hepatocytes.
胶原酶肝脏灌注后，细胞悬液经低速离心，上清液中存在小肝细胞。这些细胞在添加了10 mM烟酰胺、10%胎牛血清、1 mM抗坏血酸2 - 磷酸酯和10 ng/ml表皮生长因子的杜氏改良 Eagle 培养基中增殖超过2个月并形成集落。一个小细胞最终增殖到数百个细胞。此外，集落中的一些细胞显示分化为成熟肝细胞，这些成熟肝细胞具有大的细胞质，有时还有两个细胞核。培养过程中，肝细胞在培养基中白蛋白的分泌随时间增加，且细胞在其细胞膜中具有连接蛋白32和许多过氧化物酶体。因此，小肝细胞可能是可进一步分化为成熟肝细胞的“定向祖细胞”。
