成年小鼠肝组织中长期体外培养的探讨

目的探讨成年肝组织(AHT)体外中长期培养的各项活性指标,寻求以AHT替代成年肝细胞(AHC)培养的方法。方法以0.5mm3大小的肝组织贴壁于盖破片上,静置培养48h后改为旋转培养,每2天作半量换液1次,并取盖玻片作HE染色观察肝组织内细胞存活状况,第26天时结束培养观察;取培养第6天的肝组织设对照组、大黄素实验组、胆红素代谢组,分别行大黄素抗内毒素实验和胆红素代谢实验。结果在对照组,AHT染色鲜明、细胞存活良好;大黄素实验组呈现大黄素的抗内毒素作用而明显提高AHT的细胞活性;胆红素代谢实验组的总胆红素、直接胆红素、间接胆红素分别下降13.69%、19.85%、10.38%。结论AHT保持了体内三维立体结构,体外培养存活期可达26天,生物代谢活性良好,并有省时、操作简便、低成本的优点。

A relatively long-term adult mouse hepatic tissue culture in vitro

Objective To investigate hepatic tissue culture in vitro for a long-term in adult mice.Methods (0.5mm~3)hepatic tissue in group A were adherenced in coverslip and statically cultured for 48h,then were revolved,and the medium was half exchanged every 2 days.The survival conditions of hepatic tissue were observed by HE staining for 26 days.The antiendotoxin effect of emodin in group B was tested by MTT in the 6th day,and the value of light absorption was detected by AliSei auto-analyzer.The metabolic activity of bilirubin in group C was determined by Beckmann auto-analyzer in the 6th day.Results Staining of hepatic tissue in group A was bright and the survival condition was fine;emodin in group B showed the antiendotoxin effect and obviously elevated the cytoactive of hepatic tissue,and total bilirubin and direct bilirubin in group C decreased by 13.69% and 19.85%,respectively.Conclusion The survival time of cultured adult mouse hepatic tissue can reach 26 days in vitro,and the metabolic activity is fine.This culture method is simple and low-cost.