多细胞因子组合对人脐血单个核细胞体外扩增及粘附分子和CXCR4表达的影响

The effect of several cytokines combinations on in vitro expansion of mononuclear cells from cord blood and their expression of adhesive molecules and CXCR4

目的:探讨不同细胞因子组合对人脐血单个核细胞体外扩增及扩增后粘附分子和CXCR4表达的影响。方法:将新鲜脐血标本分离的单个核细胞接种于含有不同细胞因子组合的无血清无基质培养体系中培养7天,在0、7天检测有核细胞数(TNC)、CD34+细胞数及CD34+CXCR4+、CD34+CD49d+、CD34+CD62L+的细胞数和集落形成单位(CFU)数。根据不同细胞因子组合实验分组为:对照组;SCF+FL(简称SF)组;SFT(SCF+FL+TPO)组;SFT6(SCF+FL+TPO+IL-6)组;SFTs(SCF+FL+TPO+sIL-6R)组;SFT6s(SCF+FL+TPO+IL-6/sIL-6R)组。结果:和对照组相比,SF、SFT、SFT6、SFTs、SFT6s组均可有效地扩增脐血造血细胞(P<0.05),SFT、SFT6、SFTs、SFT6s四组扩增效果优于SF,差异有显著性(P<0.05),但SFT和SFT6、SFTs三组之间却无明显区别(P>0.05),在SFT6组合基础上加入sIL-6R后,即SFT6s组能有效地扩增脐血细胞,并优于SFT、SFT6、SFTs三组(P<0.05);SF、SFT、SFT6、SFTs四组细胞因子组合均可提高脐血CD34+细胞上CD49d、CD62L和CXCR4的表达,但四组之间差异无显著性(P>0.05),SFT6s组可明显促进脐血CD34+细胞上CD49、CD62L和CXCR4的表达,并优于SF、SFT、SFT6、SFTs四组,差异有显著性(P<0.05)。结论:IL-6/sIL-6R可协同SCF、FL和TPO有效地扩增脐血细胞并能促进和归巢有关的CD49d。

Objective：To explore the effect of different Cytokines on the expansion of the mononuclear cells drived from umbilical cord blood and the expression of adhesive molecules and of CXCR4 on the CD34^＋cells.Methods：Human fresh CB mononuclear cells were cultured in serum-free and stroma-free medium containing different combinations of cytokines for 7 days,the total cells were counted at day 0 and 7 CD34^＋ cells and CD34^＋CXCR4^＋,CD34^＋CD49d^＋ and CD34^＋CD62L^＋cells were assayed by flow cytometry.According to the different cytokine combination,six groups：the blank control;SF（SCF＋FL） ;SFT（SCF＋FL＋TPO）;SFT6（SCF＋FL＋TPO＋IL-6） group;SFTs（SCF＋FL＋TPO＋ sIL-6R） group;SFT6s（SCF ＋FL＋TPO＋IL-6/sIL-6R）,were divided.Results：SF,SFT,SFT6,SFTs,SFT6s all could effectively promote the cell peo liferation and CD34^＋ cells CFU formation in vitro.where SFT,SFT6,SFTs and SFT6s were superior to the group SF.There was no difference among group SFT,SFT6 and SFTs（P〉0.05）.The addition of sIL-6R to group SFT6 restored UCB cells expansion to a higher extent than in group SFT,SFT6 and SFTs（P〈0.05）;SF,SFT,SFT6,SFTs and SFT6s all could upregulate the expression of CD49d,CD62L and CXCR4 on the expanding CD34^＋ cells from cord blood,and the effect of SFT6s was superior to those of SF,SFT,SFT6 and SFTs（P〈0.05）,but there was no difference between SF,SFT,SFT6 and SFTs（P〉0.05）.Conclusion：The IL-6/sIl-6R in combination with SCF,FL and TPO could enhance the expansion of cord blood CD34^＋ cells and their expression of CD49d,CD62L and CXCR4.