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目的分离、培养人脑胶质瘤肿瘤浸润淋巴细胞并对其进行活性测定，探讨临床应用肿瘤浸润淋巴细胞的理论根据。方法采用机械分离、酶解聚和不连续密度梯度离心方法从１０例脑胶质瘤制备肿瘤浸润淋巴细胞单细胞悬液，体外培养扩增；采用ＮＡＧ酶荧光比色法测定肿瘤浸润淋巴细胞杀伤活性。结果肿瘤浸润淋巴细胞得率达到７２５±３２１；在ＲＰＭＩ１６４０完全培养基中生长旺盛，第３、４周能够达到１０９数量级，此时杀伤活性亦达到高峰。结论肿瘤浸润淋巴细胞适当条件体外培养扩增，在数量和杀伤活性等方面能够满足临床要求，培养第３、４周是肿瘤浸润淋巴细胞应用的时机。
Objective To isolate and culture human glioma tumor-infiltrating lymphocytes and measure their activity, and to explore the theoretical basis of clinical application of tumor-infiltrating lymphocytes. Methods Single cell suspensions of tumor-infiltrating lymphocytes were prepared from 10 gliomas by mechanical separation, enzymatic hydrolysis and discontinuous density gradient centrifugation. The single cell suspensions of tumor infiltrating lymphocytes were cultured and expanded in vitro. The cytotoxicity of tumor infiltrating lymphocytes active. Results The rate of tumor-infiltrating lymphocytes reached 725 ± 321. In RPMI1640 complete medium, the growth rate was very high. The first and third week could reach 109 orders of magnitude, and the killing activity reached a peak at this time. Conclusions Tumor infiltrating lymphocytes can be expanded in vitro under appropriate conditions and can meet the clinical requirements in terms of quantity and cytotoxic activity. The third and fourth week of culture is the timing of the application of tumor infiltrating lymphocytes.