体外培养环境对脐血CD34~+造血干/祖细胞基因表达的影响

目的研究体外培养环境对脐血CD34+造血干/祖细胞基因表达的影响。方法脐血单个核细胞(MNC)在静态和动态培养系统中培养7d后,收集CD34+造血干/祖细胞,利用SMART-PCR技术从少量RNA中扩增cDNA用于标记探针,与基因芯片杂交后分析培养前后以及不同培养模式下培养的CD34+造血干/祖细胞基因表达的差异。结果在所检测的588个基因中,45个基因在培养前后的CD34+造血干/祖细胞中差异表达,其中20个基因在培养后表达上调,25个基因表达下调。另外,12个基因在不同培养模式中差异表达,只有CCL2在动态培养中高表达,而其余11个基因均在静态培养中高表达。结论通过分析体外培养环境对CD34+造血干/祖细胞基因表达的影响,可以在分子水平上理解CD34+造血干/祖细胞对培养环境的生理响应。

Influence of Culture Environment In Vitro on Gene Expression of CD34 + Hematopoietic Stem/Progenitor Cells from Umbilical Cord Blood

Objective To study the influence of culture environment in vitro on the gene expression of CD34~ hematopoietic stem/progenitor cells from umbilical cord blood(UCB).Methods Cultivate the mononuclear cells(MNCs) from UCB in static and stirred culture systems for 7 d,then collect CD34~ hematopoietic stem/progenitor cells(HSPCs).Extract total RNA from the collected HSPCs using Trizol reagent and amplify sufficient cDNA by SMART-PCR for the labeling of probe.Screen the differentially expressed genes with Atlas cDNA arrays covering 588 genes.Results The differential expression of a total of 45 genes were found in CD34~ HSPCs before and after cultivation,of which 20 genes were up-regulated and 25 genes were down-regulated.In addition,the expression levels of 12 genes were significantly different in the CD34~ HSPCs cultivated in static and stirred systems.With the exception of CCL2,the rest genes were highly expressed in the CD34~ HSPCs in static culture.Conclusion Quantitative molecular understanding could provide new insights into the physiological responses of CD34~ HSPCs to culture environments.