CELLULAR INTERACTIONS PROMOTE TISSUE-SPECIFIC FUNCTION, BIOMATRIX DEPOSITION AND JUNCTIONAL COMMUNICATION OF PRIMARY CULTURED-HEPATOCYTES
CELLULAR INTERACTIONS PROMOTE TISSUE-SPECIFIC FUNCTION, BIOMATRIX DEPOSITION AND JUNCTIONAL COMMUNICATION OF PRIMARY CULTURED-HEPATOCYTES
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DOI：
10.1002/hep.1840080506
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发表时间：
1988-09-01
期刊：
影响因子：
13.5
通讯作者：
MORIN, O
中科院分区：
文献类型：
--
作者：
GOULET, F;NORMAND, C;MORIN, O
将从正常成年大鼠肝脏制备的肝细胞接种到胶原基质上，单独培养或在大鼠肝内皮细胞存在下培养。当肝细胞单独培养在一个明确定义的无血清培养基中，白蛋白的产生减少和快速的形态学恶化的胆小管结构和缝隙连接发生在4至5天。与此相反，肝细胞与肝间充质细胞共培养至少4周保持形态完整和生化功能。它们重组成小岛，继续分泌高水平的白蛋白，不表达α-甲胎蛋白（胎儿标记物），并保持强烈的染料偶联。所有的肝细胞合成白蛋白，并保留其缝隙连接通道。肝细胞和内皮细胞之间没有观察到连接通讯。长纤维含有纤维连接蛋白，I型胶原和层粘连蛋白分布在肝细胞诱导共培养，但从来没有出现在单独培养的肝细胞。此外，补充苯巴比妥和二甲基亚砜，这两个提高肝细胞培养的寿命和功能活动的实质性定义的培养基，未能诱导肝细胞纯培养物中的网硬蛋白纤维形成。当肝细胞与各种上皮细胞或间充质细胞[大鼠肝上皮细胞（T51 B）、小鼠胚胎成纤维细胞（NIH 3 T3）、人或大鼠真皮成纤维细胞和牛主动脉内皮细胞（AG 4762）]结合时，也可获得在与肝窦细胞培养的肝细胞中观察到的白蛋白分泌、生物基质沉积和连接通讯的调节。
Hepatocytes, prepared from normal adult rat liver, were seeded onto a collagen substratum and cultured alone or in the presence of rat liver endothelial cells. When hepatocytes were cultured alone in a hormonally defined serum-free medium, decreased albumin production and rapid morphological deterioration of bile canaliculi structures and gap junctions occurred within 4 to 5 days. In contrast, hepatocytes cocultured with liver mesenchymal cells remained morphologically intact and biochemically functional for at least 4 weeks. They reorganized into small islands, continued to secrete high levels of albumin, did not express .alpha.-fetoprotein (a fetal marker), and remained strongly dye coupled. All of the hepatocytes synthesized albumin and retained their gap junctional channels. No junctional communication was observed between hepatocytes and endothelial cells. Long fibers containing fibronectin, Type I collagen and laminin distributed over the hepatocytes were induced in coculture but never appeared in hepatocytes cultured alone. Moreover, supplementation of the hormonally defined medium with phenobarbital and dimethyl sulfoxide, both of which improve the life span and functional activities of cultured hepatocytes, failed to induce reticulin fiber formation in pure culture of hepatocytes. The modulation of albumin secretion, biomatrix deposition and junctional communication observed in hepatocytes cultured with sinusoidal liver cells was also obtained when hepatocytes were in association with various epithelial or mesenchymal cells [rat liver epithelial cells (T51B), mouse embryonic fibroblasts (NIH 3T3), human or rat dermal fibroblasts and bovine aorta endothelial cells (AG 4762)].