微囊化猪肝细胞培养的研究

In vitro culture of microencapsulated porcine hepatocytes

目的 微囊包裹猪原代肝细胞并进行普通培养 ,观察肝细胞形态变化。方法 在不含小牛血清的RPMI 164 0培养基中培养 ,光镜下观察培养过程中肝细胞形态变化 ,检测不同时期培养上清中白蛋白及尿素含量。结果 微囊肝细胞可较长期存活 ,微囊肝细胞组与游离肝细胞组上清蛋白分泌在前 3d差异无显著性 (P >0 .0 5 ) ,4d后差异有显著性 (P <0 .0 5 ) ,而上清尿素合成在前 2d差异无显著性 (P >0 .0 5 ) ,3d后差异有显著性 (P <0 .0 5 )。结论 微囊材料可制备微囊化猪原代肝细胞 ,并进行体外培养 ,与游离肝细胞比较具有更好分泌合成功能。

Objective To observe the changes in cell morphology of microencapsulated and non-microencapsulated hepatocytes in culture.Methods Microencapsulated and non-microencapsulated hepatocytes were inoculated in the culture medium containing RPMI 1640 supplemented with insulin,dexamethasone but without bovine calf serum.The morphologic changes of cultured microencapsulated and non-microencapsulated hepatocytes were observed and the concentration of albumin and urea in the supernatant in different culture periods were examined.Results There was no significant difference in the secretion of supernatant proteins in the first three days between microencapsulated hepatocytes and non-microencapsulated hepatocytes (P>0.05),but there was significant difference 4 days later (P<0.05).There was no significant difference in urea synthesis in the supernatant in the first two days between microencapsulated hepatocytes and non-microencapsulated hepatocytes (P>0.05),but there was significant difference 3 days later (P<0.05).Conclusion The microencapsulated porcine hepatocytes can retain viability and differentiating function for 14 days in vitro culture,better and longer than the non-microencapsulated.