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为了观察早期作用造血细胞因子SCF、FL、IL-3、IL-6、TPO单独及联合应用,对脐血CD34+细胞的体外扩增作用。我们用吸附单克隆抗体一磁珠分离系统富集人脐血CD34+细胞,在体外液体培养体系中加入不同的细胞因子扩增4周,每周取样计数有核细胞总数及集落形成细胞(CFC)数。结果表明:用磁性细胞分离议富集脐血CD34+细胞纯度为80%~87%;一些细胞因子有明显的协同效应,其联合应用的扩增作用显著高于单因于作用:SCF+FL存在下,IL-3是有效扩增有核细胞总数及CFC的关键因子了;细胞因子SCF+FL+IL-3和SCF+FL+IL-3+IL-6组合对有核细胞总数及CFC均有良好的扩增效应,培养2周时对CFC的扩增倍数分别为38.3±4.4和29.6±2.7倍,可满足成人移植及基因治疗等的需要。
In order to observe the early role of hematopoietic cytokines SCF, FL, IL-3, IL-6, TPO alone and in combination, on cord blood CD34 + cells in vitro expansion. We enriched human umbilical cord blood CD34 + cells by adsorbing monoclonal antibody-magnetic separation system. Different cytokines were added to the in vitro liquid culture system for amplification for 4 weeks. Total number of nucleated cells and colony forming cells (CFCs) number. The results showed that the purity of CD34 + cells isolated from cord blood by magnetic cell separation was about 80% -87%. Some cytokines had a synergistic effect, and their combined effect was significantly higher than that of SCF + FL alone. IL-3 is the key factor to effectively amplify the total number of nucleated cells and CFC. The combination of cytokines SCF + FL + IL-3 and SCF + FL + IL-3 + IL-6 has a good amplification effect on the total number of nucleated cells and CFC. CFC amplification fold was 38.3 ± 4.4 and 29.6 ± 2.7 times, respectively, to meet the needs of adult transplantation and gene therapy.