期刊
STEM CELL REPORTS
卷 7, 期 5, 页码 911-926出版社
CELL PRESS
DOI: 10.1016/j.stemcr.2016.09.012
关键词
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资金
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - Ministry of Science, ICT and Future Planning [2015R1A2A2A01003604, 2015R1A5A1009701]
- Next-Generation BioGreen 21 Program - Rural Development Administration, Republic of Korea [PJ01133802]
- National Research Foundation of Korea [2015R1A2A2A01003604] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Naive and primed pluripotent stem cells (PSCs) and germ cells express the Oct4 gene. The Oct4 gene contains two cis-regulatory elements, the distal enhancer (DE) and proximal enhancer (PE), which differentially control Oct4 expression in a cell-type-specific and stage-specific manner. Here, we generated double transgenic mice carrying both Oct4-Delta PE-GFP and Oct4-DDE-tdTomato (RFP), enabling us to simultaneously monitor the activity of DE and PE. Oct4 expression is stage-specifically regulated by DE and PE during embryonic and germ cell development. Using this dual reporter system, we successfully cultured pure populations of naive (GFP(+)RFP(-)) and primed (GFP(-)RFP(+)) PSCs. We found that GFP(+)RFP(-) cells were metastable (not naive) in serum-containing medium; stable naive pluripotent cells were observed in medium containing two inhibitors (Meki and GSKi) but lacked serum. Finally, we suggest that the activity of Oct4 DE and PE is regulated by the repressive histone marks and DNA methylation in a cell-type-specific manner.
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