Fouad F M, Scherer R, Abd-El-Fattah M, Ruhenstroth-Bauer G
Eur J Cell Biol. 1980 Jun;21(2):175-9.
Morphologically intact rat hepatocytes separated by collagenase perfusion were cultured in L-15/fetal calf serum medium to form a monolayer. Thereafter the hepatocytes were grown in serum-free L-15 medium in which they produced and continuously released plasma proteins. The secreted plasma proteins were collected, separated and characterized by crossed immunoelectrophoresis. Most of the newly biosynthesized plasma proteins secreted into the medium during incubation for thirty hours had the same electrophoretic mobility, antigenicity and staining characteristics as their counterparts in rat serum. The addition of tritium labelled amino acid mixture to the culture medium revealed that the release of radioactively labelled plasma proteins into the culture medium was essentially linear during the thirty hour incubation period. However, saturation of the intracellular pool took place after ca. ten hours of incubation. Addition of leukocytic endogenous mediator, LEM, to cultures of rat hepatocytes caused a profound increase in the relative concentration of acute-phase proteins secreted into the culture medium.
通过胶原酶灌注分离得到的形态完整的大鼠肝细胞，在L - 15/胎牛血清培养基中培养形成单层。此后，肝细胞在无血清的L - 15培养基中生长，在此培养基中它们产生并持续释放血浆蛋白。收集分泌的血浆蛋白，通过交叉免疫电泳进行分离和鉴定。在孵育30小时期间分泌到培养基中的大多数新生物合成的血浆蛋白，其电泳迁移率、抗原性和染色特性与大鼠血清中的对应物相同。向培养基中添加氚标记的氨基酸混合物表明，在30小时的孵育期内，放射性标记的血浆蛋白向培养基中的释放基本呈线性。然而，孵育约10小时后细胞内池发生饱和。向大鼠肝细胞培养物中添加白细胞内源性介质（LEM），导致分泌到培养基中的急性期蛋白的相对浓度显著增加。
