应用Solexa技术检测脐血CD34^+细胞体外诱导的巨核细胞mRNA表达  

Analysis of mRNA Expression Profiles of Megakaryocytes from Human Cord Blood CD34^+ Cells Ex Vivo Expanded Using Solexa Sequencing

作　　者：王芳[1] 何吉[1] 朱发明[1] 刘晋辉[1] 秦斐[1] 陈舒[1] 徐罡[1] 吕行军[1] 严力行[1] 

机构地区：[1]浙江省血液中心血液安全研究卫生部重点实验室,杭州310006

出　　处：《中国医学科学院学报》2011年第5期529-532,共4页Acta Academiae Medicinae Sinicae

摘　　要：目的应用Solexa技术研究人脐带血CD34+细胞体外诱导巨核细胞的mRNA表达。方法采用密度梯度离心法和免疫磁珠分选系统获取人脐带血CD34+细胞。100 ng/ml促血小板生成素诱导培养12 d后,应用抗CD41+单克隆抗体免疫磁珠法分选获得巨核细胞和非巨核细胞。应用Solexa技术检测巨核细胞和非巨核细胞的mRNA表达差异。结果获取的Tag初始数量巨核细胞和非巨核细胞分别为3 773 147和3 533 805个。整个清晰的Tag数量分别为3 291 132和2 967 947个,明显独特的tag数量分别为197 769和245 318个。其中上调基因表达数量为1161个,下调为902个。上调Tag表达数量为2717个,下调为1519个。结论巨核细胞和非巨核细胞mRNA表达存在显著差异,差异基因编码产物与细胞发育、黏附、凋亡、代谢、细胞内及细胞间的信号转导以及免疫应答等功能相关,进一步深入将有助于探讨巨核细胞的表达机制、信号传导及调控机制。Objective To investigate the mRNA expression profiles of megakaryocytes（MKs） from human cord blood CD34＋ cells ex vivo expanded using Solexa technique.Methods CD34＋ Cells were isolated using density gradient centrifugation and magnetic activated cell sorting.Cultures were stimulated with recombinant human thrombopoietin（100 ng/ml）.After 12 days,the MKs fraction was separated from the non-MKs fraction using an anti-CD41 monoclonal antibody by immunomagnetic sorting.The mRNA expression of MKs and non-MKs was detected by Solexa sequencing.Results We obtained 3773147 and 3533805 Tags from MKs and non-MKs,respectively.The amounts of unambiguous tags were 3291132 and 2967947 and those of distinct tags were 197769 and 245318.The expression of 1161 genes was up-regulated and that of 902 genes down-regulated.The expression of 2717 tags was up-regulated and that of 1519 tags down-regulated.Conclusions MKs and non-MKs have remarkably different mRNA expression profiles.The differential gene-encoded products may be involved in cellular development,adhesion,apoptosis metabolism,intra-and inter-cellular signal transduction,and immune response.Further studies on this topic may clarify the expression mechanism,signal transduction,and regulation mechanisms.

关 键 词：巨核细胞 MRNA表达 体外扩增 Solexa测序 

分 类 号：R392.12[医药卫生—免疫学]