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目的建立尼龙毛分离纯化人外周血T淋巴细胞的方法,并探讨其增殖活性。方法密度梯度离心法分离正常人外周血单个核细胞,以尼龙毛分离出T细胞,使用流式细胞仪鉴定细胞纯度、回收率;MTT法检测细胞增殖活性。结果尼龙毛法分离后T淋巴细胞纯度为(70.70±3.45)%,稍高于分离前(66.71±5.47)%(t=1.746,P>0.05)。回收率为(42.4±6.95)%,用植物血凝素(PHA)刺激分离后的T淋巴细胞的刺激指数(SI)为(1.80±0.27)。结论在细胞生物学研究中,尼龙毛分选T淋巴细胞纯度不高,PHA刺激T淋巴细胞的增殖性较强。
Objective To establish a method for the isolation and purification of human peripheral blood T lymphocytes from nylon hair and to explore the proliferative activity. Methods T-cells were isolated from normal human peripheral blood mononuclear cells by density gradient centrifugation. Purity and recovery of cells were determined by flow cytometry. Cell proliferation was measured by MTT assay. Results The purity of T lymphocytes after nylon wool separation was (70.70 ± 3.45)%, slightly higher than that before separation (66.71 ± 5.47)% (t = 1.746, P> 0.05). The recovery rate was (42.4 ± 6.95)%. The stimulation index (SI) of isolated T lymphocytes stimulated with phytohemagglutinin (PHA) was (1.80 ± 0.27). Conclusion In the study of cell biology, the purity of T lymphocytes in nylon hair sorting is not high, and PHA stimulates the proliferation of T lymphocytes.