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

DOI:
10.3791/56391-v

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11:53 min

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June 01, 2018

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, ,
1State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences

Capitoli

  • 00:04Titolo
  • 00:57Grow Mouse ES Cells on Gelatin-coated Dishes
  • 02:48Extract DNA from Cells and Enzymatic Digestion of DNA into Single Nucleosides
  • 05:29Analyze 5mC and 5hmC Using UHPLC-MS/MS
  • 08:34Results: Methylation Status of Mouse Embryonic Stem Cells
  • 10:17Conclusion

Summary

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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.

Tags

Mouse Embryonic Stem CellsGenomic HypomethylationMEK Inhibitor PD0325901Vitamin CCulture MethodDNA ExtractionDNA MethylationPluripotencyHeterogeneityMorphologyUndifferentiated State

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