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目前测定人NK活性大都用放射性同位素的方法,如~(51)Cr释放法和~(125)I-udR释放法虽有一定优越性,但由于所需试剂和仪器的限制,使普通实验室难以开展。我们设计了一种简单的测定NK活性的方法,用乳酸脱氢酶(LDH)作指标来表示NK活性,现简介如下:(一)用淋巴细胞分离液(比重1.078)常规分离周围血淋巴细胞,洗后用RPMI164C培养液(含小牛血清10％,Hepes 25mM,谷酰胺30mg％,双抗常规量,pH7.2)配成500万细胞/ml浓度的细胞悬液作为效应细胞。(二)用体外传代悬浮生长的K562细胞作靶细胞。实验前两天换一次新鲜1640培养液,实验时用培养
At present, although the methods for measuring human NK activity using radioactive isotopes, such as ~(51)Cr release method and ~(125)I-udR release method, have some advantages, due to limitations of required reagents and instruments, the general laboratory Difficult to carry out. We have designed a simple method to measure NK activity, using lactic acid dehydrogenase (LDH) as an indicator to express NK activity, as follows: (i) Routine isolation of peripheral blood lymphocytes using lymphocyte separation fluid (specific gravity 1.078) After washing, a cell suspension with a concentration of 5 million cells/ml was prepared as effector cells using RPMI164C culture medium (containing calf serum 10%, Hepes 25 mM, glutamine 30 mg%, double-antibody conventional amount, pH 7.2). (B) In vitro suspension of K562 cells grown in suspension as target cells. Change the fresh 1640 culture solution two days before the experiment and use the culture during the experiment.