鸡离体肝细胞几种分离方法的比较

Comparison of Several Methods for Isolating Chicken Hepatocytes

对鸡肝细胞离体培养中几种分离肝细胞的方法进行了比较 .试验选用 1～ 5 6日龄的粤黄鸡 ,分别采用肝组织块培养、肝组织直接通过细胞筛过滤、2 .5g/L胰蛋白酶消化、2 .5g/L胰蛋白酶加 0 .1g/LEDTA消化、不同时间的4℃冷消化与 37℃消化、淋巴细胞分离液分离、5g/L氯化铵除去红细胞等试验方法 ,观察了鸡离体肝细胞的分离及培养效果 .结果表明 :采用 2 5g/L胰蛋白酶加 0 .1g/LEDTA消化 10min、用淋巴细胞分离液分离后的肝细胞培养效果最好 ,肝细胞在培养液中分裂增殖 。

Comparison was made among several methods for isolating chicken hepatocytes. Yuehuang chicken of 1～56 days old were sacrificed and hepatocytes cultivated from bits of liver tissue, dispersed cell prepared by passing through a cell sieve, digestion with 2 5 g/L pepsin, and digestion with 2 5 g/L pepsin plus 0 1 g/L EDTA. The digestion temperature was 37℃and 4℃. In order to get rid of erythrocytes inside the medium, the reagent for isolating lymphocytes and 5 g/L ammonium chloride were used. The resultes showed that high density and viability of chicken hepatocytes could be obtained after digestion with 2 5 g/L pepsin plus 0 1 g/L EDTA at 37℃ for 10 minutes.