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背景和目的:造血干细胞具有自我更新、多向分化与重建长期造血的潜能,因此,造血干细胞广泛应用于干细胞移植、免疫治疗、基因治疗等领域。胎儿脐带血中富含造血干细胞,但单份脐带血中造血干细胞的数量有限,不能充分满足临床和科研需要,因此在体外培养使脐带血干/祖细胞数量扩增的研究日益受到重视。已知一些细胞因子可以在体外培养中使脐血干/祖细胞大量扩增。新近发现IL-6/sIL-6R(可溶性IL-6受体)或其融合蛋白可促使脐带血CD34+细胞中CD34+gp130+IL-6R-细胞亚群在体外培养中大量扩增。本实验旨在观察IL-6/sIL-6R在脐带血CD34+细胞体外扩增中的作用,并探讨合适细胞因子组合。方法:脐血CD34+细胞用MiniMACS分离,然后在含有不同细胞因子组合的液体培养基中体外培养7天或14天,培养前后分别进行有核细胞计数、用FCM(流式细胞术)测定CD34+细胞比例计算CD34+细胞总数及进行CFU-GM集落培养。根据不同细胞因子组合分为空白对照组,SCF组,SCF+IL-6/sIL-6R组,SCF+FL+IL-6/sIL-6R组,SCF+FL组。结果:空白对照组和SCF组CD34+细胞数量在培养7天或14天后明显下降。SCF+IL-6/sIL-6R组培养7、14天分别使有核细胞及CD34+细胞绝对数扩增(7.1±2.4)倍、(39.0±14.0)倍及(1.8±0.7)倍、(4.8±2.4)倍;SCF+FL+IL-6/sIL-6R组(16.5
BACKGROUND & OBJECTIVE: Hematopoietic stem cells have the potential of self-renewal, multidirectional differentiation and reconstruction of long-term hematopoiesis. Therefore, hematopoietic stem cells are widely used in the fields of stem cell transplantation, immunotherapy and gene therapy. Fetal umbilical cord blood is rich in hematopoietic stem cells. However, the quantity of hematopoietic stem cells in single umbilical cord blood is limited and can not meet the needs of clinical and scientific research. Therefore, the research on the number expansion of umbilical cord blood stem / progenitor cells in vitro has received more and more attention. It is known that some cytokines can greatly expand cord blood stem / progenitor cells in vitro culture. Recently, it has been found that IL-6 / sIL-6R (soluble IL-6 receptor) or its fusion protein can promote large-scale expansion of CD34 + gp130 + IL-6R- cell subpopulation in cord blood CD34 + cells in vitro culture. The aim of this study was to investigate the role of IL-6 / sIL-6R in the in vitro expansion of cord blood CD34 + cells and to explore suitable combinations of cytokines. Methods: Umbilical cord blood CD34 + cells were isolated by MiniMACS and then cultured in liquid medium containing different combinations of cytokines for 7 days or 14 days. The nucleated cells were counted before and after culture. CD34 + cells were determined by FCM (flow cytometry) Proportions were calculated for the total number of CD34 + cells and CFU-GM colonies were cultured. According to different combinations of cytokines, they were divided into blank control group, SCF group, SCF + IL-6 / sIL-6R group, SCF + FL + IL-6 / sIL-6R group and SCF + FL group. Results: The number of CD34 + cells in blank control group and SCF group decreased significantly after 7 days or 14 days. The absolute numbers of nucleated cells and CD34 + cells were (7.1 ± 2.4) fold, (39.0 ± 14.0) and (1.8 ± 0.7) fold and (4.8 ± 0.7) fold respectively in SCF + IL-6 / ± 2.4) fold; SCF + FL + IL-6 / sIL-6R group (16.5