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目的 初步探讨粘附NK细胞的制备方法及免疫学特性。方法 首先经淋巴细胞分离液、贴壁粘附及尼龙毛柱粘附对外周血NK细胞进行初步分离 ,然后联合使用Percoll不连续密度梯度离心法和T细胞Panning法进一步纯化NK细胞 ,应用流式细胞仪鉴定NK细胞并检测其纯度。rhIL 2 (6 0 0 0IU ml)诱导纯化的NK细胞 (R NK)转变为粘附NK(A NK)细胞 ,然后利用细胞计数法、MTT法和RT PCR方法检测A NK细胞的增殖和杀伤等免疫学特性。结果 经过纯化、诱导获得较高纯度、较高活性的A NK细胞。结论 初步建立了利用人外周血NK细胞制备A NK细胞的方法
Objective To investigate the preparation methods and immunological characteristics of adherent NK cells. Methods Peripheral blood NK cells were preliminarily separated by lymphocyte separation, adherent adhesion and nylon wool column adhesion. Then NK cells were further purified by Percoll discontinuous density gradient centrifugation and T cell Panning method. The flow cytometry Cytometer identifies NK cells and detects their purity. rhIL 2 (60 000 ml) induced NK cells (NK cells) to become adherent to NK (A NK) cells, and then assayed for proliferation and killing of A NK cells by MTT and RT PCR Immunological characteristics. The results of purification, induced higher purity, higher activity of A NK cells. Conclusion A method of preparing A NK cells using human peripheral blood NK cells was initially established