Identification of mesenchymal stem cell (MSC)-transcription factors by microarray and knockdown analyses, and signature molecule-marked MSC in bone marrow by immunohistochemistry
Identification of mesenchymal stem cell (MSC)-transcription factors by microarray and knockdown analyses, and signature molecule-marked MSC in bone marrow by immunohistochemistry
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DOI：
10.1111/j.1365-2443.2009.01281.x
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发表时间：
2009-03-01
期刊：
影响因子：
2.1
通讯作者：
Kato, Yukio
中科院分区：
文献类型：
--
作者：
Kubo, Hiroshi;Shimizu, Masakazu;Kato, Yukio
虽然体外扩增间充质干细胞（MSC）已被用于许多研究，但MSC的分子特征和体内分布状况仍不清楚。为了解决这个问题，我们使用DNA微阵列和实时RT-PCR分析鉴定了许多人类msc特征基因，包括9个转录因子基因：大多数msc特征基因在与成骨、软骨或脂肪诱导培养基孵育24小时后，或在9个转录因子敲除48-72小时后下调。此外，ETV1、ETV5、FOXP1、GATA6、HMGA2、SIM2或SOX11的下调抑制了MSC的自我更新能力，而FOXP1、SOX11、ETV1、SIM2或PRDM16的下调则降低了成骨和/或成脂潜能。此外，利用针对间充质干细胞特征分子（包括GATA6、TRPC4、FLG和tgm2）的抗体进行免疫组化，发现成年小鼠的内皮附近和骨髓内部存在间充质干细胞样细胞。这些结果表明，MSC在体内和体外都能合成一组MSC标记物，并且MSC特征转录因子参与了MSC干性调控。
Although ex vivo expanded mesenchymal stem cells (MSC) have been used in numerous studies, the molecular signature and in vivo distribution status of MSC remain unknown. To address this matter, we identified numerous human MSC-characteristic genes-including nine transcription factor genes -using DNA microarray and real-time RT-PCR analyses: Most of the MSC-characteristic genes were down-regulated 24 h after incubation with osteogenesis-, chondrogenesis- or adipogenesis-induction medium, or 48-72 h after knockdown of the nine transcription factors. Furthermore, knockdowns of ETV1, ETV5, FOXP1, GATA6, HMGA2, SIM2 or SOX11 suppressed the self-renewal capacity of MSC, whereas those of FOXP1, SOX11, ETV1, SIM2 or PRDM16 reduced the osteogenic- and/or adipogenic potential. In addition, immunohistochemistry using antibodies for the MSC characteristic molecules-including GATA6, TRPC4, FLG and TGM2-revealed that MSC-like cells were present near the endosteum and in the interior of bone marrow of adult mice. These findings indicate that MSC synthesize a set of MSC markers in vitro and in vivo, and that MSC-characteristic transcription factors are involved in MSC stemness regulation.