Mitaka T, Norioka K, Sattler G L, Pitot H C, Mochizuki Y
Department of Pathology, Sapporo Medical University, School of Medicine, Japan.
Cancer Res. 1993 Jul 1;53(13):3145-8.
The proliferation of primary cultured rat hepatocytes was observed in serum-free modified Dulbecco's modified Eagle's medium supplemented with 10 mM nicotinamide and 10 ng/ml of epidermal growth factor. These proliferating cells were mainly mononucleate and formed small-cell colonies after Day 4. The small cells in focal colonies were surrounded by typical hepatocytes and were stained immunocytochemically with anti-rat albumin and anti-cytokeratin 8 antibodies. This suggests that the cells in the small-cell colonies were derived from hepatocytes. The frequency of appearance of small-cell colonies with age was examined by the use of primary cultured hepatocytes isolated from the livers of rats between the ages of 3 wk and 90 wk. In the cells from 4- to 5-wk-old rats, about 58 colonies per 1000 attached cells appeared 144 h after plating; the number of colonies rapidly decreased to about 25 in 6- to 8-wk-old rats. In adult rats, about 17 colonies were seen, and only about five colonies were observed in rats more than 80 wk old.
在添加了10 mM烟酰胺和10 ng/ml表皮生长因子的无血清改良杜尔贝科改良伊格尔培养基中观察到原代培养大鼠肝细胞的增殖。这些增殖细胞主要为单核细胞，在第4天后形成小细胞集落。局灶性集落中的小细胞被典型的肝细胞包围，并用抗大鼠白蛋白和抗细胞角蛋白8抗体进行免疫细胞化学染色。这表明小细胞集落中的细胞来源于肝细胞。通过使用从3周龄至90周龄大鼠肝脏分离的原代培养肝细胞，研究了小细胞集落随年龄出现的频率。在4至5周龄大鼠的细胞中，接种后144小时，每1000个贴壁细胞中约出现58个集落；在6至8周龄大鼠中，集落数量迅速减少至约25个。在成年大鼠中，约可见17个集落，而在80周龄以上的大鼠中仅观察到约5个集落。
