摘要:
目的探讨改良、简化肝细胞的灌注、分离方法,以提高细胞产量,降低制作成本.方法对传统的两步原位胶原酶灌流法进行改进,改变灌注方式,分离的大鼠肝细胞采用三次低速离心,台盼兰染色检测活率,并与Percoll梯度离心纯化法进行比较.结果大鼠肝细胞经低速离心后,活率仅(48.38±6.97)%.三次离心后再经Percoll梯度离心纯化后活率为(92.63±2.50)%,纯度>95%,纯化前后活率比较差异有显著性(P<0.01).结论该方法要求设备简单、容易操作,细胞产量、活率及纯度高.
Abstract:
Objective To establish an improved,simplified and effective hepatocyte isolation technique in order to increase cell production and decrease the prime cost.Methods The improved simple perfusion system was used to isolate rat hepatocytes by a two-step perfusion with collagenase in situ.Hepatocytes were separated from the dissociative cells by low speed centrifugation(500 rpm,3 min,3 times),and discontinous gradient centrifugation method was applied to purify the hepatocytes.The hepatocyte yields and viabilit...
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