目的改进一种简易、高效的乳小鼠肝细胞培养法,并对所培养的细胞进行盘定。方法原代培养采用改进的组织块贴壁方法,以少量培养基孵育1~2h。传代培养采用0.25%胰蛋白酶消化随后以小牛血清培养为单层细胞。整个过程无需离心。结果成功地用这种简便的方法获得了纯度较高的肝细胞并进行了传代和鉴定。结论使用此方法培养肝细胞简单、高效、快捷,适合大多数实验室培养非大规模肝细胞。
Objective To search for an easier and higher efficiency method for the primary culture of murine hepatoeytes,and identify it. Methods The method of tissue piece adherence was improved for the primary culture. The harvested hepatocytes were kept in a humidified incubator with a little medium about 1 - 2 h. The serial subcultivation of routine monolayer hepatocytes was performed in a simple way, in which hepatocytes were digested by 0.25 % trypsin and cultured with calf serum medium. The whole procedure did not require eentrifugation. Results The higher purity hepatoeytes and its passage cells was sueeessfuUy obtained. For further identifing this method, the primary culture of murine hepatocytes was performed all suecessfuUy many times. Conclusion The method for the primary culture of murine hepatocytes is a good and economic method for the general lab to culture the hepatocytes.