大鼠α-谷氨酰转肽酶阳性肝细胞在无血清培养液中的生长

THE GROWTH OF RAT γ -GLUTAMYLTRANSPEPTIDASE- POSITIVE HEPATOCYTES IN SERUM-FREE MEDIUM

原位肝胶原酶灌流分离出经化学致癌剂诱发的大鼠癌前期肝细胞,用“淘洗”技术纯化γ-谷氨酰转肽酶(GGT)阳性肝细胞,置无血清的DMEM:F12(1:1)培养液中培养。结果表明,GGT阳性或阴性肝细胞均不能在此培养液中长期生长。而经乙基亚硝基脲(ENU)处理4h的肝细胞,只有GGT阳性肝细胞能在此无血清培养液中长期生长并传代,其形态及排列类似正常肝细胞,电镜下可见典型的连接复合体和毛细胆管,但核较大;组织化学及免疫组织化学方法证实这些细胞仍维持GGT阳性,且能产生白蛋白,故此种细胞可能是介于正常肝细胞与肝癌细胞的癌前期细胞。

Using in situ perfusion of collagenase, the preneoplastic hepatocytea wesisolated from rat liver -which had been subjected to chemical hepatocarcinoger|6-sis. The γ-glutamyltranspeptidase (GOT) positive hepatocytes were purified by means of the 'panning' technique and cultured in serum-free DMEM-F12 (1:1) medium. The results showed that both GGT-positive and GGT-negative hepatocytes could not survive and proliferate in this medium for a long period of time. However, GGT-positive hepatocytes which had been treated with ethylnitrosou-rea(ENU) in vitro could be subcultured in serum-free medium and multiply continuously. The morphology and arrangement of these cells were similar to normal hepatocytes, typical junctional complex (tight junction, intermediate junction and desmosome) and bile canaliculi could be seen under electromicroscopy. Moreover, these hepatocytes were revealed by immunohistocheinical and histo-chemical methods to be able to produce albumin and to keep GOT activity. Therefore, these hepatocytes may be thought to be the preneoplastic hepatocytes being between normal and neOplastic hepatocytes.