Yoda Y, Kawakami Z, Shibuya A, Abe T
Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan.
Exp Hematol. 1988 Sep;16(8):712-7.
Human bone marrow (BM) cells, depleted of nylon wool-adherent cells, T cells, and natural killer (NK) cells, were cultured in medium containing recombinant interleukin 2 (rIL2). After 21 or 24 days in culture, numerous lymphoid cells with multiple azurophilic granules and a morphology similar to large granular lymphocytes (LGL) were found. Two-color analysis of surface phenotype showed many of these cells to be NKH1-positive and a limited number of cells had other NK markers such as CD16, CD2, or CD8. The CD3 antigen was not coexpressed with NKH1. The cultured BM cells were cytotoxic for K562, Daudi, and Raji cell lines. The NKH1+, CD2-, CD3-, CD16- cells were sorted and, in addition to having the LGL morphology, were found to be cytotoxic for K562 cells (NK [K562]). The generation of NK(K562) activity was significantly suppressed by 5-bromodeoxyuridine plus ultraviolet light treatment, indicating that DNA synthesis is required. These experiments suggest that the described culture conditions allow differentiation of progenitor cells, into immature, but functionally active, NK cells.
去除尼龙毛黏附细胞、T细胞和自然杀伤(NK)细胞后的人骨髓(BM)细胞,在含有重组白细胞介素2(rIL2)的培养基中培养。培养21天或24天后,发现了许多具有多个嗜天青颗粒且形态类似于大颗粒淋巴细胞(LGL)的淋巴细胞。表面表型的双色分析显示,这些细胞中有许多为NKH1阳性,少数细胞具有其他NK标志物,如CD16、CD2或CD8。CD3抗原不与NKH1共表达。培养的BM细胞对K562、Daudi和Raji细胞系具有细胞毒性。对NKH1 +、CD2 -、CD3 -、CD16 -细胞进行分选,发现这些细胞除具有LGL形态外,对K562细胞(NK [K562])也具有细胞毒性。5-溴脱氧尿苷加紫外线处理可显著抑制NK(K562)活性的产生,表明DNA合成是必需的。这些实验表明,所述培养条件可使祖细胞分化为未成熟但功能活跃的NK细胞。