Zhang Jing, Zhu Jing-Song, Li Qian-Ru, DU Ying, Yang Bo, Li Guo-Xi, Hu Xiang
Department of Microbiology and Immunology, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450001, Henan Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Feb;18(1):146-50.
This study was aimed to investigate the changes of differentiation and gene expression of CD133(+) cells in human umbilical cord blood induced by stem cell factor (SCF) and basic fibroblast growth factor (bFGF) in vitro, and to explore the biological characteristics of CD133(+) cells so as to provide experimental basis for potential use in regenerative medicine. The human umbilical cord blood CD133(+) cells were isolated from umbilical cord blood and purified by MACS magnetic selection. The CD133(+) cells were cultured in DMEM/F12 medium containing SCF, bFGF and B27 for 10 to 15 days. The total RNA of these cells was extracted before and after culture, and the analysis of related gene expression levels of these cells was performed using oligonucleotide microarrays. The results showed that out of 263 genes 21 genes were obviously up-regulated after culture than that before culture, whereas 7 genes were found to be significant down-regulated as compared with fresh-separated CD133(+) cells. These genes were involved in stem cell specific markers, cell cycle regulators, stem cell differentiation markers and signaling pathways that are important for stem cell maintenance. It is concluded that SCF and bFGF can induce differentiation of human cord blood CD133(+) cells through up- or down-regulation of specific genes. This study provides gene expression information for SCF and bFGF-induced human cord blood CD133(+) cells and contributes to understand the effect of SCF and bFGF on proliferation and differentiation CD133(+) cells at gene level, and promotes therapeutic applications of the CD133(+) cells induced by SCF and bFGF.
本研究旨在探讨干细胞因子（SCF）和碱性成纤维细胞生长因子（bFGF）体外诱导人脐血CD133(+)细胞分化及基因表达的变化，探索CD133(+)细胞的生物学特性，为其在再生医学中的潜在应用提供实验依据。从脐血中分离出人脐血CD133(+)细胞，并通过MACS磁选法进行纯化。将CD133(+)细胞在含有SCF、bFGF和B27的DMEM/F12培养基中培养10至15天。培养前后提取这些细胞的总RNA，使用寡核苷酸微阵列对这些细胞的相关基因表达水平进行分析。结果显示，在263个基因中，有21个基因在培养后比培养前明显上调，而与新鲜分离的CD133(+)细胞相比，有7个基因显著下调。这些基因涉及干细胞特异性标志物、细胞周期调节因子、干细胞分化标志物以及对干细胞维持至关重要的信号通路。结论是，SCF和bFGF可通过上调或下调特定基因来诱导人脐血CD133(+)细胞分化。本研究提供了SCF和bFGF诱导人脐血CD133(+)细胞的基因表达信息，有助于从基因水平了解SCF和bFGF对CD133(+)细胞增殖和分化的影响，并促进SCF和bFGF诱导的CD133(+)细胞的治疗应用。
