摘要
为了获得人脐血造血干细胞(HSCs)的microRNAs(miRNAs)表达谱,并对相关miRNAs功能进行初步鉴定.利用免疫磁珠(MACS)和流式细胞仪(FACS)细胞分选技术分离人脐血造血干细胞(HSCs),分别提取细胞总RNA并分离小分子RNA,经荧光标记后与miRNAs基因芯片杂交,获得HSCs的miRNAs表达谱,集落形成实验(CFC)研究在HSC中高表达miR-520h对HSC的促分化作用.成功分离人脐血CD34+细胞和HSC,经基因芯片杂交获得31个造血干细胞相关miRNAs,其中22个为低表达,9个为高表达;经实时定量RT-PCR验证miR-520h显著升高,CFC实验表明其可增加多种集落形成,具有促进HSC向祖细胞分化的作用.上述结果表明,人脐血HSC具有自身特征性miRNAs,参与并调控HSC生物学功能,为深入探讨miRNAs在造血系统发育中的作用打下基础.
In order to obtain MicroRNA （miRNAs） expression profiles of human hematopoietic stem cells （HSCs）, and to preliminarily investigate functions of HSC-correlative miRNAs, by using miniMACS magnetic beads and fluorescence-activated cell sorting （FACS）, isolated hematopoietic stem cells （HSCs） were isolated from human umbilical cord blood and performed total RNA extraction. Next, using a microarray, miRNA gene expression profiles of HSCs were obtained. CFC assays were performed to research on differentiation-promoting function of miR-520h, enriched in HSCs. Results showed that CD34^＋ hematopoietic cells and HSCs were successfully isolated from human umbilical cord blood, and 31 HSCs-correlative miRNAs were screened by microarrray. Among them, 22 were low in HSCs, and 9 were high. The result of real time RT-PCR confirmed high expression of miR-520h in HSCs. CFC assays showed that miR-520h promotes differentiation of HSCs into progenitor cells. In conclusion, human HSCs have a set of specific miRNAs that contribute to regulation of HSCs functions, which pave the way for exploring the roles of miRNAs in development of hematopoiectic system.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2007年第11期1190-1196,共7页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金项目(30400241
30672076)
中国博士后科学基金资助项目(2005038478).~~
