不同粘附剂对大鼠肝实质细胞体外培养的影响

Effection of Different Adhesives on the Culture of Rat Hepatocytes

目的:比较鼠尾胶与多聚赖氨酸对大鼠肝实质细胞体外培养的影响。方法:分别采用鼠尾胶与多聚赖氨酸包被同一块培养板,然后将从大鼠肝脏中分离出来的肝实质细胞,接种到包被好的培养板中。于接种前(0 h),接种后体外培养24 h、72 h显微镜下观察细胞贴壁与形态变化情况。结果:接种前(0 h)可见新鲜分离的肝实质细胞呈圆形,明亮,有立体感,轮廓完整,层次清楚;体外培养24 h后两种粘附剂包被的同一块培养板中,均可观察到肝实质细胞正常生长,且细胞形态由圆形转变为多角形,并且融合聚集,胞体变平整,贴壁情况区别不大;培养72 h后细胞间开始出现连接,大部分肝细胞呈现出双核或多核,并且多聚赖氨酸包被的培养板中可见大量肝细胞呈岛屿状,已完全贴壁于培养板上。结论:多聚赖氨酸作为包被材料更有利于肝实质细胞贴壁生长以及保持细胞固有形态。

Objective:Comparison of the effects of rat tail’s collagen and polylysine on the culture of rat hepatocytes in vitro.Methods:The same culture plate was coated with rat tail’s collagen and polylysine respectively,and then the hepatocytes isolated from rat liver were inoculated into the coated culture plate.The changes of cell adhesion and morphology were observed under microscope before inoculation(0 h)and after inoculation for 24 hours and 72 hours in vitro.Results:Before inculation(0 h),the freshly isolated hepatocytes were round,bright,three-dimensional,complete in outline and clean in layer.After 24 hours of culture in vitro,the normal growth of hepatocytes were observed in the same culture plates coated with two kinds of adhesives,and the morphology of hepatocytes changed from round to polygonal,and the cells fused and gathered,the cell body became flat,and there was little difference in adhesion.After 72 hours of culture,the connection between the cells began to appear.Most of the hepatocytes were binuclear or multinucleated,and a large number of hepatocytes were island-shaped in the polylysine-coated culture plate,which had completely adhered to the culture plate.Conclusions:As a coating material,polylysine is more beneficial to the adherent growth of hepatocytes and maintain the inherent morphology of hepatocytes.