摘要
为探讨乳猪肝细胞分离、体外培养的条件及其生长方式。方法用两步灌注结合胶原酶消化法分离乳猪肝细胞，用激素条件培养基进行体外培养，ＥＤＴＡ消化传代，测定细胞的活性，观察其生长方式，群体倍增时间及蛋白质和ＤＮＡ的合成。结果在胶原酶浓度为０．０７５％，细胞的收获和成活率较高，原代和传代的肝细胞生长良好，原代肝细胞倍增时间较短，ＤＮＡ和蛋白质的合成也较活跃，与传代细胞相比，无明显差别（Ｐ＞０．０５）。结论乳猪肝细胞分离所需的胶原酶浓度较高，在一定的条件下，能分裂增殖，且能进行传代，同时能维持其基本特性。
Aims In an attempt to use pup pig hepatocgtes as the cellular element of hybridiZation bioartificial liver.Methods hepatocytes were isolated from pup pigs by pond vein cannulation and the two step perfusion methods combined withcollagenase type Ⅳ digestion, then cultured via conditioned medium. The growth Pattern, population doubling time, the synthesisof DNA of the hepatocytes were investigated in the Primary and Passaged cultures by conventional methed. Results It was foundto get higher harvest and viability of hepatocytes, the concentration of collagenase type Ⅳ should be at 0.075% .The growth Pattern of the two cultrs were similar, but the population doubling time in primary culture was shorter than that in the Passaged,the synthesis of DNA and Protein in the Primary was also more vigores than that in the Passaged. Conclusions The isolation ofpup pig hepatocytes requires higher concentlation of collagenase type Ⅳ; in certain conditions, it could be culted and Passagedfor a long time, meanwhile maintaining its characteristics of proliferation.
出处
《胃肠病学和肝病学杂志》
CAS
1999年第1期20-22,共3页
Chinese Journal of Gastroenterology and Hepatology
