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Biochemistry

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JoVE Journal Biochemistry
An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
 

An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C

Article DOI: 10.3791/56391-v • 11:53 min • June 1st, 2018
June 1st, 2018 •
1, 1, 1
1State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences

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résumé
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We described in detail two chemical-based protocols for culturing mouse embryonic stem cells. This new method utilizes synergistic mechanisms of promoting Tet-mediated oxidation (by vitamin C) and repressing de novo synthesis of 5-methylcytosine (by PD0325901) to maintain DNA hypomethylation and pluripotency of mouse embryonic stem cells.

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Alternative Culture Method Genomic Hypomethylation Mouse Embryonic Stem Cells MEK Inhibitor PD0325901 Vitamin C Pluripotent State FBS Cultured Mouse Embryonic Stem Cells Morphology Dehypermethylation Excellent Morphology Undifferentiated State Plates Buffers Prewarm Culture Medium Trypsin PBS Passage The Cells Trypsin EDTA Serum Medium Single Cell Suspension Gelatin-coated Dish
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