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作者通过含重组白细胞介素2(rIL-2)基质中培养事先清除成熟T细胞和自然杀伤细胞(NK)的筛选骨髓细胞,证实NK来源骨髓。取自6名健康志愿者胸骨的肝素化骨髓细胞,经Ficoll-Hypaquè(FH)离心分离单个核细胞,使混于含5%小牛血清的RPMI-1640培养液,置尼龙羊毛柱管37℃孵育1小时,滤出柱管中不粘附细胞(NW-NA)。取其1.5×10~7个细胞,加入经氢溴2氨甲基异巯基脲处理的绵羊红细胞1.5×10~9个,作FH离心分离。取洗出的界面非花结形成细胞加于1μg/ml多种单克隆抗体(T101、OKM1、OKTl l、Leullb)混合液中,4℃孵育45分钟。最后用无毒性兔补体37℃孵育45分钟,离心成为筛选骨髓细胞。后者90%以上为粒或红细胞,淋巴细胞多小至中等大,嗜阿尼林颗粒淋巴细胞罕见,无残存NKHl~+细
The authors identified NK-derived bone marrow cells by screening bone marrow cells prior to culture of mature interleukin 2 (rIL-2) stroma in advance to clear mature T cells and natural killer cells (NK). Heparinized bone marrow cells from six healthy volunteers were isolated from the mononuclear cells by Ficoll-Hypaquè (FH) centrifugation and mixed with RPMI-1640 medium containing 5% fetal bovine serum at 37 ℃ After incubation for 1 hour, non-adherent cells (NW-NA) were filtered out. Take 1.5 × 10 ~ 7 cells, adding 1.5 × 10 ~ 9 sheep erythrocytes treated with hydrobromide 2 aminomethyl isothiocarbamide to make FH centrifugal separation. The eluted interface non-flowering cells were added to 1μg / ml mixed solution of multiple monoclonal antibodies (T101, OKM1, OKT11, Leullb) and incubated for 45 minutes at 4 ° C. Finally, non-toxic rabbit complement was incubated for 45 minutes at 37 ° C, and then centrifuged to select bone marrow cells. More than 90% of the latter are granulocytes or erythrocytes, with small to medium lymphocytes and rare anilin granules, with no residual NKH1 +