Gene expression profile of primary human CD34+CD381o cells differentiating along the megakaryocyte lineage
Gene expression profile of primary human CD34+CD381o cells differentiating along the megakaryocyte lineage
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DOI：
10.1016/j.exphem.2004.04.002
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发表时间：
2004-07-01
影响因子：
2.6
通讯作者：
Reems, JA
中科院分区：
文献类型：
--
作者：
Shim, MH;Hoover, A;Reems, JA
目标。为了鉴定参与巨核生成的基因，高密度寡核苷酸微阵列被用于比较未分化的CD34(+)CD38（lo）细胞和培养来源的巨核细胞（mk）的转录谱。材料与方法。将原代人CD34(+)CD38（lo）细胞培养于含白细胞介素-3、白细胞介素-6、干细胞因子和血小板生成素的培养基中，培养10天，实现巨核细胞的体外分化。利用从三种不同器官供体中分离的造血细胞和高密度寡核苷酸微阵列进行了三次重复微阵列实验。对基因阵列数据进行分析，有304个差异表达基因（P≤0.001，倍数变化大于等于3）。已知25个高度上调的基因中有三分之一参与止血（z = 6.75），在下调的基因中没有已知与mk相关的基因。我们还在粘附和受体活性（85%）和信号转导活性（68%）的基因本体类别中发现了很大比例的上调转录本。同时，70%的转录因子功能基因下调。验证性研究表明，阵列结果与原代mks mRNA和蛋白表达水平相关。这项研究提供了人类mk的全球表达谱和一系列新的和以前未表征的候选基因，这些基因是巨核形成的重要组成部分。(C) 2004国际实验血液学学会。Elsevier Inc.出版。
Objective. To identify genes involved in megakaryopoiesis, high-density oligonucleotide microarrays were used to compare transcript profiles from undifferentiated CD34(+)CD38(lo) cells and culture-derived megakaryocytes (MKs).Materials and Methods. Megakaryocyte differentiation was achieved in vitro by inducing primary human CD34(+)CD38(lo) cells in serum-deprived media supplemented with the cytokine combination of interleukin-3, interleukin-6, stem cell factor, and thrombopoietin for 10 days. Three replicate microarray experiments were performed using hematopoietic cells isolated from three different organ donors and high-density oligonucleotide microarrays.Results. Analysis of gene array data resulted in 304 differentially expressed genes (P less than or equal to 0.001, fold change greater than or equal to3). A third of the 25 most highly up-regulated genes were known to participate in hemostasis (z = 6.75), and no genes known to be associated with MKs were among the down-regulated genes. We also found a large proportion of up-regulated transcripts in gene ontology categories of adhesion and receptor activity (85%) and signal transduction activity (68%). At the same time, 70% of genes within transcription factor functions were down-regulated. Confirmatory studies indicated that the array results correlated with mRNA and protein expression levels in primary MKs.Conclusion. This study provides a global expression profile of human MKs and a list of novel and previously uncharacterized candidate genes that are important components of megakaryopoiesis. (C) 2004 International Society for Experimental Hematology. Published by Elsevier Inc.