抗TGF-β1单克隆抗体对脐血CD34^+细胞体外扩增作用的研究  

Effects of anti-TGF-β1 Monoclonal Antibody on in vitro Expansion of Cord Blood CD34^+ Cells

作　　者：栗宇[1] 刘复强[1] 王景文[1] 

机构地区：[1]首都医科大学附属北京同仁医院血液内科,北京100730

出　　处：《中国实验血液学杂志》2009年第6期1546-1550,共5页Journal of Experimental Hematology

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。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 demonstra-ted 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 eviden

关 键 词：TGF-1单克隆抗体 脐血 c—kit CD34+细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]