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采用抗CD3或抗CD8单克隆抗体的补体细胞毒方法清除脐血单个核细胞 (MNC )中T淋巴细胞 ,CD34免疫亲和柱纯化MNC中CD34 +细胞 ,流式细胞技术 (FACS )分析细胞表面标志。将CD34 +细胞中加入含多种造血生长因子的培养基进行体外液态扩增 ,并观察粒 巨噬集落 (CFU GM )和多向祖细胞集落 (CFU GEMM )形成能力。结果CD3+或CD8+细胞清除组和MNC经CD34免疫亲和柱纯化后 ,CD34 +细胞分别为 5 9 5 2 %、 5 6 70 %和 5 0 72 % ,比未纯化组 (1 0 7% )纯度大幅度提高。抗CD3单抗清除 +CD34纯化组和单纯CD34纯化组经造血生长因子刺激培养第 14天 ,细胞总数分别扩增 110 40倍和 87 0 0倍。抗CD3单抗清除 +CD34纯化组、抗CD8单抗清除 +CD34纯化组和单纯CD34纯化组的CFU GM产率分别为 2 86 5 0± 12 0 2、2 88 5 0± 17 68和 2 19 5 0± 5 3 0 3,前者虽高于后者 ,但差异无显著性。CFU GEMM产率分别为 376 67± 43 2 4、 438 33± 36 73和 311 0 0± 40 11,抗CD3单抗清除 +CD34纯化组无显著差异 ,抗CD8单抗清除 +CD34纯化组显示出显著性作用 (P <0 0 5 )。
T lymphocytes from cord blood mononuclear cells (MNC) were depleted by complement cytotoxicity using anti-CD3 or anti-CD8 monoclonal antibodies. CD34 + cells were purified by CD34 immunoaffinity column and analyzed by flow cytometry (FACS) Sign. The CD34 + cells were cultured in liquid medium containing various hematopoietic growth factors, and the formation ability of CFU GM and CFU GEMM was observed. Results The percentage of CD34 + cells in CD3 + or CD8 + cell depletion group and MNC group after purification by CD34 immunoaffinity column was 59.5%, 56.7% and 50.22% respectively, which was significantly higher than that of non-purified group (107%) substantial improvement. On the 14th day after anti-CD3 monoclonal antibody depletion + CD34 purification group and pure CD34 purification group were stimulated by hematopoietic growth factor, the total number of cells were amplified by 110 40 times and 87 0 0 times respectively. The CFU GM yields of anti-CD3 McAb + CD34 purified group, anti-CD8 mAb + CD34 purified group and CD34 purified group were respectively 2 86 5 0 ± 12 0 2 88 5 0 ± 17 68 and 2 19 5 0 ± 5 3 0 3, the former is higher than the latter, but the difference was not significant. The CFU GEMM yields were 376 67 ± 43 2 4, 438 33 ± 36 73, and 311 0 0 ± 40 11, respectively. There was no significant difference between anti-CD3 mAb + CD34 purification group and anti-CD8 mAb + CD34 purification group Significant effect (P <0 05).