胎儿骨髓间质干细胞与细胞因子对脐血CD34^+细胞扩增作用

The effect of fetal bone marrow mesenchymal stem cells and cytokines on CD34 positive cells proliferation

目的 探讨胎儿骨髓间质干细胞对CD34+ 细胞体外扩增的造血支持作用。方法 体外分离、纯化胎儿骨髓间质干细胞 ;Mini MACS免疫磁珠分选 3份脐血CD34+ 细胞 ;建立胎儿骨髓间质干细胞与CD34+ 细胞共培养体系 :第 1组为单独CD34+细胞培养 ,第 2组为胎儿骨髓间质干细胞 +CD34+ 细胞共培养 ,第 3组为细胞因子 (干细胞因子 ,白细胞介素 3,Flt3配体 ,血小板生成素 ) +CD34+ 细胞共培养 ,第 4组为胎儿骨髓间质干细胞 +细胞因子 +CD34+ 细胞共培养。用流式细胞仪检测不同培养时间的CD34+ 细胞。结果 胎儿骨髓间质干细胞表达CD2 9,CD4 4 ;免疫磁珠分选CD34+ 细胞的平均纯度为 97.4 % ;胎儿骨髓间质干细胞 +CD34+ 细胞共培养 2 8d ,有核CD34+ 细胞仍占有核细胞的 6 .4 3% ;CD34+ 细胞在胎儿骨髓间质干细胞、细胞因子作用下培养 2 8d ,有核细胞总数、CD34+ 细胞数分别被扩增 1.6 5× 10 5倍、788倍。结论 胎儿骨髓间质干细胞可有效扩增脐血造血干 /祖细胞。

Objective To elucidate the role of CD34+ cells expansion supported by fetal mesenchymal stem cells (MSCs) in vitro.Methods Fetal MSCs were isolated and purified in vitro.Cord blood CD34+ cells were isolated by using Mini-MACS separation system.The experiments included four groups,i.e.,1.only CD34+ cells, 2.MSCs and CD34+ cells,3.cytokines:stem cell factor (SCF);interleukin-3(IL-3);Flt3-ligand(FL);thrombopoeitin (TPO) and CD34+ cells,and 4.MSCs cytokines and CD34+ cells.The percentage of CD34+ cells derived from different cultured time was detected by flow cytometer.Results Fetal MSCs expressed CD29 and CD44.The average purity of CD34+ cells was 97.4%.After culture of 28 days,6.43% of CD34+ cells remained in the first group,the amplification of the total cells and CD34+ cells reached 1.65×105-fold and 788-fold increase in the fourth group, respectively.Conclusions Fetal MSCs can support efficiently proliferation of hematopoietic stem/progenitor cells derived from cord blood.