腫瘤藥物敏感性試驗（1）－－腫瘤細胞分離與培養

Tumor Chemosensitivity Assay:Ⅰ.Isolation and Cultivation of Tumor Cells

腫瘤體外藥物敏感性試驗已廣泛用以測定腫瘤細胞對抗腫瘤藥物的敏感性。本法現已用於篩選抗腫瘤新藥，或預測腫瘤患者對化學治療的反應。從腫瘤標本分離培養腫瘤細胞，是進行腫瘤藥物敏感性試驗的第一步，也是最重要的步驟之一。在本文中，我們報告從手術切除的實質性腫瘤標本，分離、培養、鑑定與純化腫瘤細胞的方法。腫瘤細胞係用酵素分離法進行分離（預滲序列分離法），應用梯次多皿培養法接種，用含10%胎牛血清的RPMI-1640培養液培養；通過形態學和免疫細胞化學研究進行鑑定，才能做腫瘤藥物敏感性試驗。在2.5年的時間內，我們共做52例實質性腫瘤標本，包括乳癌23例、結直腸癌17例、胃癌5例及其他腫瘤7例。其中39例（75%）能獲得較滿意的腫瘤細胞培養，可以進行腫瘤藥物敏感性試驗。在技術較成熟後2年內的37例中，細胞分離培養成功，而能做腫瘤藥物敏感性試驗的比例達到81%（30/37），效果尚稱滿意。

In vitro tumor chemosensitivity assay has been widely used to determine the response of tumor cells to anticancer drugs. It has been used for screening new anticancer drugs or for predicting the response of tumor to chemotherapeutic agents in individual patients. The first step and one of the most important steps for performing the chemosensitivity assay is to isolate and establish tumor cell cultures from the tumor specimens.We reported the methods used for isolation, cultivation, identification and purification of tumor cells from surgical specimens of various solid tumors. Tumor cells were isolated by enzyme disaggregation method (pre-penetration sequence method), seeded by sequent multi-dish culture method and cultured with RPMI-1640 medium supplemented with 10% fetal bovine serum. Tumor cells were detached after confluence of primary culture, identified by morphological and immunocytochemical studies before performing the chemosensitivity assay. During the past 2.5 years, 52 cases of various solid tumors were treated, including 23 cases of breast cancer; 17 cases of colorectal cancer, 5 cases of gastric cancer and 7 cases of other tumors. Tumor cell cultures were established in 39 cases so that the chemosensitivity assay could be performed (75%). After 6 months practice, better results were obtained with an assay evaluability rates of 81% was achieved in 30 out of 37 cases.