抗转化生长因子β1单克隆抗体对脐血CD34(+)细胞体外扩增的影响

[Effects of anti-TGF-beta1 monoclonal antibody on in vitro expansion of cord blood CD34(+) cells].

作者信息

Li Yu, Liu Fu-Qiang, Wang Jing-Wen

机构信息

Department of Hematology, Beijing Tongren Hospital, Capital University of Medical Sciences, Beijing 100730, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2009 Dec;17(6):1546-50.

PMID:20030944

Abstract

In order to investigate the effect of anti-TGF-beta1 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-beta1 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) x 10(5), (1.16 +/- 0.29) x 10(5), (1.09 +/- 0.26) x 10(5)] was significantly higher than that in control group [(1.25 +/- 0.13) x 10(5), (0.55 +/- 0.19) x 10(5), (0.51 +/- 0.2) x 10(5)](p < 0.01). The expansion of more primitive CD34(+)c-kit(-) subpopulation in test group [(12.95 +/- 3.17) x 10(3)] was even significantly higher than in control group (1.71 +/- 0.83) x 10(3) (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) x 10(3), (60.5 +/- 20.96) x 10(3)] was higher than that in control group [(5.0 +/- 2.58) x 10(3), (16.25 +/- 7.93) x 10(3)] (p < 0.01). The number of relatively mature CFU-GM between test group and control group was not statistical significance [(6.33 +/- 2.85) x 10(3) vs (4.0 +/- 2.28) x 10(3)](p > 0.05), but both were higher than that in blank group (0.75 +/- 0.29) x 10(3). These results demonstrated that anti-TGF-beta1 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-beta1 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.

摘要

为研究抗转化生长因子β1（TGF-β1）单克隆抗体对脐血CD34(+)细胞扩增的影响，将纯化的脐血CD34(+)细胞分为三组：空白对照组：第0天培养的纯化脐血CD34(+)细胞；对照组：在含干细胞因子（SCF）、白细胞介素-3（IL-3）、白细胞介素-6（IL-6）和FMS样酪氨酸激酶3配体（FLT3-L）的培养体系中培养3天的细胞；试验组：在与对照组相同的培养体系中培养3天，但添加抗TGF-β1单克隆抗体的细胞。检测三组的单个核细胞计数（MNC）、CD34和c-kit的表达以及粒-红-巨核-巨噬细胞集落形成单位（CFU-GEMM）、爆式红系集落形成单位（BFU-E）和粒-巨噬细胞集落形成单位（CFU-GM）计数。结果显示，试验组MNC、CD34(+)细胞和CD34(+)c-kit(+)细胞的扩增倍数[（2.35±0.25）×10⁵、（1.16±0.29）×10⁵、（1.09±0.26）×10⁵]显著高于对照组[（1.25±0.13）×10⁵、（0.55±0.19）×10⁵、（0.51±0.2）×10⁵]（p<0.01）。试验组中更原始的CD34(+)c-kit(-)亚群的扩增倍数[（12.95±3.17）×10³]甚至显著高于对照组（1.71±0.83）×10³（p<0.01）。集落形成试验表明，试验组中更早期祖细胞集落CFU-GEMM、BFU-E的数量[（16.3±4.72）×10³、（60.5±20.96）×10³]高于对照组[（5.0±2.58）×10³、（16.25±7.93）×10³]（p<0.01）。试验组与对照组之间相对成熟的CFU-GM数量无统计学差异[（6.33±2.85）×10³对（4.0±2.28）×10³]（p>0.05），但两者均高于空白组（0.75±0.29）×10³。这些结果表明，抗TGF-β1单克隆抗体促进了MNC和CD34(+)细胞的扩增，更显著地促进了更原始祖细胞——CD34(+)c-kit(-)细胞的扩增。同时，它增加了更不成熟集落CFU-GEMM和BFU-E的产出，但对成熟髓系集落CFU-GM没有明显影响。结论是，抗TGF-β1单克隆抗体可与其他细胞因子协同作用，有效增强脐血CD34(+)祖细胞的增殖，更重要的是能保留更多原始祖细胞。