Kralovánszky J, Harrington F, Greenwell A, Melnick R
National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
In Vivo. 1990 May-Jun;4(3):201-4.
The application of the collagenase portal vein perfusion technique for the isolation of intestinal cells resulted in the preparation of highly viable enterocytes. Cell viability was found to be greater than 90% as tested by LDH release and Trypan blue exclusion techniques. According to the results of marker enzyme determinations, collected cells were mostly of matured villus type, characterized by high disaccharidase and very low thymidine kinase activity. In vitro treatment of the isolated cells with the anticancer agent cis-diamminedichloroplatinum (II) caused decrease of the metabolic processes, i.e. glucose oxidation and protein synthesis, demonstrating that beyond the production of DNA-crosslinks other mechanisms may play a role in the cytotoxic effect of the drug. It should be stressed, however, that prolonged incubation of the cell suspension over 30 min at physiological temperature may itself lead to gradual decrease of the viability and to disturbance of the metabolic activity of the cells.
应用胶原酶门静脉灌注技术分离肠道细胞，成功制备出高活力的肠上皮细胞。通过乳酸脱氢酶释放法和台盼蓝排斥法检测发现，细胞活力大于90%。根据标记酶测定结果，收集到的细胞大多为成熟的绒毛型，其特征是二糖酶活性高，胸苷激酶活性极低。用抗癌药物顺二氯二氨铂（II）对分离出的细胞进行体外处理，导致代谢过程即葡萄糖氧化和蛋白质合成减少，这表明除了产生DNA交联外，其他机制可能在该药物的细胞毒性作用中发挥作用。然而，应该强调的是，细胞悬液在生理温度下长时间孵育超过30分钟本身可能会导致活力逐渐下降以及细胞代谢活性受到干扰。
