Experimental Study Of The Effect On Biological Characteristics Of Hepatocyte That Co-cultured With Hepatic Stellate Cells On Microcarriers

Objective To study the biological characteristics of hepatocyte that co-cultured with hepatic stellate cells on microcarriers,and explore the effect on biological charac- teristics of the hepatocyte.Mathods The SD rat hepatocytes obtained by modified way of semi-in collagenase digestion,and then co-cultured with the rHSC-99 hepatic stellate cell line on Cytodex-3. To examine the function of hepatocytes regularly and observe the morphlogical change of hepatocytes by reverse microscopy continuously during the cultivation.Results During the process of cultivation,the number of activative cells, the albumin synthesis, urea production and diazepam transformation of hepatocytes of all group, are increase firstly and decrease subsequently. There is a top on 4th day. From 4th day to 14th day, the albumin synthesis, urea production and diazepam transformation of hepatocytes co-cultured with hepatic stellate cells on cytodex-3(Group D) is hither than that co-cultured with hepatic stellate cells on TCPs(Group B) and mono-cultured on cytodex-3(Group C)(p<0.05); and that of the Group of hepatocytes mono-cultured on TCPs(Group A) is always lower than Group B, Group C and Group D (p<0.05)from 2nd day to 14th day. The 4th day's total RNA of all Group is on a high level, and that of Group D is higher than Group B, that of Group C is higher than Group A. The morphological characteristics of hepatocytes maintain 10 days in group A,but more than 2 weeks in Group B and C; Part of hapetcytes in group D can maintain more than 3 weeks.Conclusion The characteristical function of hepatcytes cultured with hepatic stellate cells on cytodex-3 is better than that of monocultured on cytodex-3,and better that of co-cultured on TCPs; Co-cultured with hepatocytes and hepatic stellate cells on microcarriers, is an ideal way to culture hepatcytes in vitro.