RRC ID |
50087
|
著者 |
Yamamoto T, Takenaka C, Yoda Y, Oshima Y, Kagawa K, Miyajima H, Sasaki T, Kawamata S.
|
タイトル |
Differentiation potential of Pluripotent Stem Cells correlates to the level of CHD7.
|
ジャーナル |
Sci Rep
|
Abstract |
Embryonic Stem Cells (ESC) possesses two distinct features; self-renewal and the potential to differentiate. Here we show the differentiation potential and growth rate of ESC correlates positively with the expression level of the gene encoding chromodomain helicase DNA binding protein 7 (CHD7). When ESCs are maintained in feeder-free conditions and single cell seeding, ESC KhES-1 having 4520 copies or more of CHD7 in 5 ng total RNA show differentiation potential, but this is lost when the CHD7 copy number is reduced in KhES-1 to less than 696 by alternative culture conditions. Introduction of siCHD7 reduced differentiation potential and growth rate of KhES-1. Interestingly, KhES-1 underwent spontaneous differentiation when mCHD7 was introduced and we could not obtain CHD7-overexpressing ESC in culture. These data suggest that CHD7 drives differentiation, and there is a lower limit for CHD7 to initiate differentiation and an upper limit for CHD7 if maintained in undifferentiated state, and such upper limit varies depending on culture condition. As CHD7 drives cell growth, ESC with the highest permissible CHD7 level in the given culture become dominant in a couple of passages. Thus, we can select differentiation resistance-free cell clones by optimizing the culture system using CHD7 as an index.
|
巻・号 |
8(1)
|
ページ |
241
|
公開日 |
2018-1-10
|
DOI |
10.1038/s41598-017-18439-y
|
PII |
10.1038/s41598-017-18439-y
|
PMID |
29321579
|
PMC |
PMC5762636
|
MeSH |
Cell Culture Techniques
Cell Differentiation / genetics*
Cell Line
Cell Proliferation
Cells, Cultured
DNA Helicases / genetics*
DNA Helicases / metabolism
DNA-Binding Proteins / genetics*
DNA-Binding Proteins / metabolism
Embryonic Stem Cells / cytology
Embryonic Stem Cells / metabolism
Gene Expression Regulation, Developmental*
Humans
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / metabolism*
Protein Transport
RNA, Messenger / genetics
RNA, Messenger / metabolism
RNA, Small Interfering / genetics
|
IF |
3.998
|
引用数 |
8
|
リソース情報 |
ヒト・動物細胞 |
KhES-1(HES0001)
201B7(HPS0063) |