Identification of mesenchymal stem cell (MSC)-transcription factors by microarray and knockdown analyses, and signature molecule-marked MSC in bone marrow by immunohistochemistry.
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.
Kubo
Hiroshi
H
Department of Dental and Medical Biochemistry, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Shimizu
Masakazu
M
Taya
Yuji
Y
Kawamoto
Takeshi
T
Michida
Masahiko
M
Kaneko
Emi
E
Igarashi
Akira
A
Nishimura
Masahiro
M
Segoshi
Kazumi
K
Shimazu
Yoshihito
Y
Tsuji
Koichiro
K
Aoba
Takaaki
T
Kato
Yukio
Y
D005347:Fibroblasts / Q000166:cytology; D000091344:Filaggrin Proteins; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012867:Skin / Q000166:cytology; D014157:Transcription Factors / Q000235:genetics*
C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D014157:Transcription Factors