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目的 :观察CD34免疫亲合柱对脐血造血干 祖细胞分离纯化的效果及纯化后CD34+ 细胞的增殖分化特性。方法 :采用CD34免疫亲合柱分离脐血单个核细胞 (MNC)中的CD34+ 细胞 ,流式细胞技术 (FACS)进行细胞表面标志测定。将分离前后的细胞加入造血生长因子进行液态扩增培养和多向祖细胞集落 (CFU GEMM)培养。结果 :经CD34免疫亲合柱分离后脐血中CD34+ 细胞为 49 6 2 %± 17 6 9% ,明显高于脐血MNC(1 17%± 0 6 8% ) ,细胞回收率为 5 4 38%± 11 91%。分离后CD34+ 细胞和脐血MNC经造血生长因子刺激培养 2 0d分别扩增 5 6 1 0 0倍和 44 44倍。培养至 12d时 ,免疫亲合柱分离组CD34+ 细胞阳性率为 5 3 38% ,对照组为 7 91%。分离组CFU GEMM产率明显高于对照组 (P <0 0 0 1)。结论 :CD34免疫亲合柱应用于脐血造血干 祖细胞的分离可充分富集CD34+ 细胞 ,且分离后的CD34+ 细胞具有明显的增殖效应和CFU GEMM形成能力。
OBJECTIVE: To observe the effect of CD34 immunoaffinity column on the isolation and purification of hematopoietic stem / progenitor cells from cord blood and the proliferation and differentiation of CD34 + cells. Methods: CD34 + cells in cord blood mononuclear cells (MNCs) were isolated by CD34 immunoaffinity column. The cell surface markers were determined by flow cytometry (FACS). The cells before and after isolation were added into hematopoietic growth factors for liquid expansion culture and CFU-GEMM culture. Results: The percentage of CD34 + cells in cord blood was 49 6 2% ± 17 6 9%, significantly higher than that of cord blood MNC (1 17% ± 0 68%) after isolation by CD34 immunoaffinity column. The cell recovery was 5 4 38 % ± 11 91%. After isolated CD34 + cells and cord blood MNC by hematopoietic growth factor stimulated culture 20d were amplified 5 6 1 0 0 times and 44 44 times. At 12 days after culture, the positive rate of CD34 + cells in the immunoaffinity column was 53 38% and in the control group was 71 91%. The CFU GEMM yield of the isolated group was significantly higher than that of the control group (P <0.01). Conclusion: The CD34 immunoaffinity column can be used to enrich CD34 + cells for the isolation of cord blood hematopoietic stem and progenitor cells, and the isolated CD34 + cells have obvious proliferative effects and CFU GEMM formation ability.