差异显示法分析不同生长环境中造血干/祖细胞基因表达的研究

Gene Expression Analysis of CD34^+ Hematopoietic Stem and Progenitor Cells Grown in Different Culture Environments Using Differential Display

目的对比分析不同生长环境中的脐血CD34+造血干/祖细胞基因表达变化。方法采用静态和动态培养系统培养脐血单个核细胞,1周后收获CD34+造血干/祖细胞,提取总RNA,用差异显示法对比分析在不同生长环境中造血干/祖细胞基因表达的差异。结果在所使用的差异显示条件下,得到30个差异表达基因片段,其中一个差异表达片段为RAN基因,该基因属于RAS癌基因家族,可能与造血细胞增殖有关。结论不同生长环境影响CD34+造血干/祖细胞的基因表达,这些差异表达的基因可为优化体外培养环境,扩增造血细胞提供分子基础。

Objective： To investigate the changes of gene expression in CD34 ^＋ hematopoietic stem and progenitor cells （HSPCs） under different growth environments. Methods： Umbilical cord blood mononuclear ceils （ UCB MNCs） were cultured in static and stirred systems. After 7 days of culture, CD34 ^＋ ceils were isolated and total RNA was extracted. Gene expression patterns of CD34^＋ ceils from fresh, static and stirred cultures were compared using differential display （DD）. Results： 30 gene fragments displayed differential expression levels based on the conditions of DD. One of differentially expressed genes was identified as RAN, which is a member of oncogene RAS family. This gene may be associated with proliferation of hematopoietic ceils. Conclusion： Different growth environments induced differential gene expression patterns of CD34 ^＋ HSPCs. These differentially expressed genes would give new insights into optimizing in vitro environments for expanding hematopoietic ceils