Gebhardt R, Schrode W, Eisenmann-Tappe I
Physiologisch-chemisches Institut der Universität, Tübingen, Germany.
Cell Biol Toxicol. 1998 Feb;14(1):55-67. doi: 10.1023/a:1007468521927.
Four epithelial cell lines established from juvenile rat liver and selected on the basis of their capacity to prolong the lifespan of cocultured hepatocytes were compared with respect to several immunocytochemical markers (vimentin, cytokeratin 19, MAB 19C6), enzyme activities, and amino acid uptake systems. Their phenotypes were found to be quite different from that of hepatocytes and bile duct epithelial cells (BEC), but very similar among each other. In particular, a variety of functions affected by dexamethasone (DEX) or changing spontaneously in cultured hepatocytes and/or BEC, showed neither inducibility nor spontaneous changes in the four cell lines. Instead, the lines were inducible for glutamine synthetase (GS) by DEX, in contrast to hepatocytes and BEC but also to other juvenile or adult epithelial lines that did not support cocultured hepatocytes. In addition, they showed relatively high basal levels of GS activity, exceeding those found in adult epithelial cell lines and approaching the average values found for liver tissue. Basal as well as DEX-induced GS activity was reduced in the presence of newborn calf serum, while only DEX-induced but not basal activity was suppressed by glutamine. These results suggest an origin of these four juvenile epithelial cell lines different from that of hepatocytes as well as of BEC. Furthermore, they suggest the coherent acquisition of new functional properties during early phases of cultivation of these cell lines; the selective inducibility of GS by DEX and its suppression by glutamine are the most intriguing of these, because neither is found in any normal cell type present in rat liver.
从幼年大鼠肝脏建立的四种上皮细胞系,基于其延长共培养肝细胞寿命的能力进行筛选,对其几种免疫细胞化学标志物(波形蛋白、细胞角蛋白19、单克隆抗体19C6)、酶活性和氨基酸摄取系统进行了比较。发现它们的表型与肝细胞和胆管上皮细胞(BEC)有很大不同,但彼此之间非常相似。特别是,受地塞米松(DEX)影响或在培养的肝细胞和/或BEC中自发变化的多种功能,在这四种细胞系中既未表现出诱导性也未出现自发变化。相反,与肝细胞和BEC不同,也与不支持共培养肝细胞的其他幼年或成年上皮细胞系不同,DEX可诱导这些细胞系中的谷氨酰胺合成酶(GS)。此外,它们显示出相对较高的GS基础活性水平,超过成年上皮细胞系中的水平,接近肝脏组织中的平均值。在新生小牛血清存在下,基础以及DEX诱导的GS活性均降低,而谷氨酰胺仅抑制DEX诱导的活性,不抑制基础活性。这些结果表明这四种幼年上皮细胞系的起源与肝细胞和BEC不同。此外,它们表明在这些细胞系培养的早期阶段一致地获得了新的功能特性;DEX对GS的选择性诱导及其被谷氨酰胺抑制是其中最引人关注的,因为在大鼠肝脏中存在的任何正常细胞类型中都未发现这种情况。