一种分离自体肿瘤细胞与TIL细胞新方法的建立

New approach for lsolating tumor cells and tumor Infiltrating lymphocytes

目的 :寻求有效地从恶性胸腔积液和腹水中分离出较多的TIL细胞并保持其高度的增殖能力与抗瘤活性的方法。方法 :应用非连续密度梯度离心法与贴壁法对恶性胸腹水中的TIL进行分离 ,而后进行诱导培养 ,比较 2种方法下TIL细胞的得率、增殖速度、杀瘤活性及培养前后的表型变化。结果 :贴壁法能够获得更多的TIL细胞 ,经统计学处理差异显著 ,P <0 0 5 ;培养后 2种方法所得TIL细胞的增殖速率、杀瘤活性及表型是一致的 ,经统计学处理差异无统计学意义 ,P >0 0 5。结论 :贴壁法具有操作简便、细胞得率高的优点 ,值得临床推广应用。

OBJECTIVE:To set up an efficient way to separate much more functional tumor infiltrating lymphocyte (TIL) from malignant pleura fluid and ascites. METHODS:TILs were isolated by two ways, i.e. the discontinuous density gradient centrifugation and the new attachment method established by authors.TILs were cultivated in the same conditions, and their proliferation as well as the killing activity and the phenotype changes were detected. RESULTS:The quantity of TILs was obviously higher by the new attachment method compared with that by the way of discontinuous density gradient centrifugation,and there were no differences in the proliferation or the killing activity in vitro as well as the phenotype changes between the two ways. CONCLUSION: The way of attachment is simple and efficient for cells separation,and it is deserved to be broadly applied.