Identification of endoglin as a functional marker that defines long-term repopulating hematopoietic stem cells
Identification of endoglin as a functional marker that defines long-term repopulating hematopoietic stem cells
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DOI：
10.1073/pnas.202614899
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发表时间：
2002-11-26
影响因子：
11.1
通讯作者：
Lodish, HF
中科院分区：
文献类型：
--
作者：
Chen, CZ;Li, M;Lodish, HF
我们描述了一种策略，以获得高度富集的长期再生（LTR）造血干细胞（HSC）从骨髓侧群（SIP）细胞通过使用转基因报告基因驱动的干细胞增强子。为了分析罕见HSC群体的基因表达谱，我们开发了一种称为"恒定比率PCR"的扩增方案，其中样品和对照cDNA在同一PCR中扩增。该方案允许我们通过寡核苷酸微阵列分析使用少至Img的总RNA来鉴定在富集的LTR-HSC群体中差异表达的基因。内皮糖蛋白是一种辅助性转化生长因子13受体，在富集的HSC中差异表达。重要的是，内皮糖蛋白阳性细胞，占总SIP细胞的20%，包含所有的LTR-HSC的活动在骨髓SP。我们的研究结果表明，内皮糖蛋白，在血管生成和造血中起着重要作用，是一个功能性标志物，定义LTR HSC。我们的整体策略可能适用于其他组织特异性干细胞的标记物的鉴定。
We describe a strategy to obtain highly enriched long-term repopulating (LTR) hematopoietic stem cells (HSCs) from bone marrow side-population (SIP) cells by using a transgenic reporter gene driven by a stem cell enhancer. To analyze the gene-expression profile of the rare HSC population, we developed an amplification protocol termed "constant-ratio PCR," in which sample and control cDNAs are amplified in the same PCR. This protocol allowed us to identify genes differentially expressed in the enriched LTR-HSC population by oligonucleotide microarray analysis using as little as 1 mg of total RNA. Endoglin, an ancillary transforming growth factor 13 receptor, was differentially expressed by the enriched HSCs. Importantly, endoglin-positive cells, which account for 20% of total SIP cells, contain all the LTR-HSC activity within bone marrow SP. Our results demonstrate that endoglin, which plays important roles in angiogenesis and hematopoiesis, is a functional marker that defines LTR HSCs. Our overall strategy may be applicable for the identification of markers for other tissue-specific stem cells.