摘要
目的 探讨一种简单获取大量高活性肝细胞的培养方法。方法 用半原位酶消化技术对SD大鼠肝细胞进行分离与微载体黏附培养 ,连续观察肝细胞的形态。检测肝细胞的白蛋白分泌功能和葡萄糖合成功能 ,并同时与贴壁培养的肝细胞比较。结果 黏附培养的肝细胞存活率、白蛋白分泌功能、葡萄糖合成功能都保持较高水平 ,优于贴壁培养的肝细胞。结论 微载体培养能提供长时间保持高活性、高密度生长的肝细胞 ,为肝细胞移植 ,肝病防治以及生物人工肝等领域的研究和广泛应用提供了基础。
Objective To study a simple culturing method for obtaining hepatocytes at high cell density and high activity.Methods Adult Sprague Dewley rats'( n =5) hepatocytes were harvested and isolated by a modified enzymatic isolation technique, called semi situ collagenase digestion technique. The harvested hepatocytes were cultured in a conditioned medium on microcarriers cytodex 3 with the restriction attachment , regularly twirled and growth factors were added into it. Parallel cultures of plated hepatocytes were also conducted. Morphology of the hepatocytes was examined dynamically. The functions of albumin secretion and glucose syntheses in hepatocytes were measured.Results The cultured lymphocytes on microcarriers had a higher survival rate, function of albumin secretion and glucose syntheses, which are superior to those of plated hepatocytes.Conclusions The microcarrier's culture method can provide more liver cells, which have a higher viability and better function, for hepatocyte transplantation, liver disease therapy and bioartificial liver.
出处
《中华器官移植杂志》
CAS
CSCD
2000年第2期106-108,共3页
Chinese Journal of Organ Transplantation
