Kubo Hiroshi, Shimizu Masakazu, Taya Yuji, Kawamoto Takeshi, Michida Masahiko, Kaneko Emi, Igarashi Akira, Nishimura Masahiro, Segoshi Kazumi, Shimazu Yoshihito, Tsuji Koichiro, Aoba Takaaki, Kato Yukio
Department of Dental and Medical Biochemistry, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Genes Cells. 2009 Mar;14(3):407-24. doi: 10.1111/j.1365-2443.2009.01281.x. Epub 2009 Feb 17.
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.
尽管体外扩增的间充质干细胞（MSC）已在众多研究中得到应用，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，我们通过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：在与成骨、软骨生成或脂肪生成诱导培养基孵育24小时后，或在敲低9个转录因子48 - 72小时后，大多数MSC特征基因表达下调。此外，敲低ETV1、ETV5、FOXP1、GATA6、HMGA2、SIM2或SOX11会抑制MSC的自我更新能力，而敲低FOXP1、SOX11、ETV1、SIM2或PRDM16会降低成骨和/或脂肪生成潜力。另外，使用针对MSC特征分子（包括GATA6、TRPC4、FLG和TGM2）的抗体进行免疫组织化学分析显示，成年小鼠的骨内膜附近和骨髓内部存在类似MSC的细胞。这些发现表明，MSC在体外和体内合成一组MSC标志物，并且MSC特征转录因子参与MSC干性调节。
