【摘要】 为了探讨TGF-β1单克隆抗体对脐血CD34+细胞的扩增作用,本研究将分离纯化的脐血CD34+细胞分为3组:①空白对照组:当天分选的新鲜脐血CD34+细胞;②对照组:含SCF、FLT3-L、IL-3、IL-6四种因子组合的无血清液体培养体系中培养3天的脐血CD34+细胞;③实验组:条件同对照组,但加入了TGF-β1单克隆抗体。3组均检测单个核细胞(MNC)计数,流式细胞术检测CD34和c-kit,计数混合集落(CFU-GEMM)、红系爆式集落(BFU-E)、粒系集落(CFU-GM)。结果显示:实验组MNC、CD34+细胞、CD34+c-kit+细胞计数分别为[(2.35±0.25)×105、(1.16±0.29)×105、(1.09±0.26)×105],明显高于对照组[(1.25±0.13)×105、(0.55±0.19)×105、(0.51±0.2)×105],(p均<0.01)。实验组CD34+c-kit-亚群计数为(12.95±3.17)×103,明显高于对照组(1.71±0.83)×103,二者相比有显著性差异(p<0.01)。实验组中早期集落CFU-GEMM、BFU-E的产率[(16.3±4.72)×103,(65.0±20.96)×103]明显高于对照组[(5.0±2.58)×103,(16.25±7.93)×103](p<0.01),相对较晚期集落CFU-GM的产率在对照组[(4.0±2.28)×103]和实验组[(6.33±2.85)×103]均高于空白组[(0.75±0.29)×103],但在对照组和实验组之间无显著性差异(p>0.05)。此结果表明,TGF-β1单克隆抗体促进了脐血MNC和CD34+细胞的扩增,且早期细胞CD34+c-kit-细胞扩增更为突出;提高了早期集落CFU-GEMM和BFU-E的产量,而对相对较晚期的髓系集落CFU-GM的产量无明显影响。结论:TGF-β1单克隆抗体能协同其它生长因子有效扩增脐血CD34+细胞,并保留一定量更早期的造血祖细胞,减轻了造血祖细胞分化的压力。更多还原

【Abstract】 In order to investigate the effect of anti-TGF-β1 monoclonal antibody on the expansion of cord blood CD34+ cells,the purified cord blood CD34+ cells were divided into three groups: blank control group: purified cord blood CD34+ cells cultured on day 0;control group: cells cultured for 3 days in the culture system,containing SCF,IL-3,IL-6 and FLT3-L;test group: cells cultured for 3 days in the same culture system as control group,but with anti-TGF-β1 monoclonal antibody. The mononuclear cell counting (MNC),the expression of CD34 and c-kit,and CFU-GEMM,BFU-E and CFU-GM counting were detected in all three groups. The result showed that the expansion of MNCs,CD34+ cells and CD34+c-kit+ cells in test group [(2.35±0.25)×105,(1.16±0.29)×105,(1.09±0.26)×105] was significantly higher than that in control group [(1.25±0.13)×105,(0.55±0.19)×105,(0.51±0.2)×105](p<0.01). The expansion of more primitive CD34+c-kit-subpopulation in test group [(12.95±3.17)×103] was even significantly higher than in control group (1.71±0.83)×103 (p<0.01). Colony forming assay showed that the number of earlier progenitor colony CFU-GEMM,BFU-E in test group [(16.3±4.72)×103,(60.5±20.96)×103] was higher than that in control group [(5.0±2.58)×103,(16.25±7.93)×103] (p<0.01). The number of relatively mature CFU-GM between test group and control group was not statistical significance [(6.33±2.85)×103 vs (4.0±2.28)×103](p>0.05),but both were higher than that in blank group (0.75±0.29)×103. These results demonstrated that anti-TGF-β1 monoclonal antibody promoted the expansion of MNC and CD34+ cells,and even more marked expansion of the more primitive progenitor cells-CD34+c-kit-cells. Meanwhile,it enhanced the output of more immature colony CFU-GEMM and BFU-E,but had no evident influence on the mature myeloid colony CFU-GM. It is concluded that the anti-TGF-β1 monoclonal antibody can synergize other cytokines to enhance the proliferation of cord blood CD34+ progenitor cells effectively,and it is more important that can reserve more primitive progenitor cells.更多还原