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Derivation and characterization of mouse embryonic stem cells from permissive and nonpermissive strains.

Anne Czechanski | Candice Byers | Ian Greenstein | Nadine Schrode | Leah Rae Donahue | Anna-Katerina Hadjantonakis | Laura G Reinholdt
Nature protocols | 2014

Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbred mouse strains. Recently, there have been considerable advances in the field of embryonic stem cell biology, particularly in the area of pluripotency maintenance in the epiblast from which the mESCs are derived. Here we describe a protocol for efficient derivation of germline-competent mESCs from any mouse strain, including strains previously deemed nonpermissive. We provide a protocol that is generally applicable to most inbred strains, as well as a variant for nonpermissive strains. By using this protocol, mESCs can be derived in 3 weeks and fully characterized after an additional 12 weeks, at efficiencies as high as 90% and in any strain background.

Pubmed ID: 24504480

Research resources used in this publication

None found

Antibodies used in this publication

None found

Associated grants

  • Agency: NIH HHS, United States
    Id: U42-OD010921
  • Agency: NCI NIH HHS, United States
    Id: P30 CA034196
  • Agency: NIH HHS, United States
    Id: U42-OD011102
  • Agency: NICHD NIH HHS, United States
    Id: R01-HD052115
  • Agency: NICHD NIH HHS, United States
    Id: R01 HD052115
  • Agency: NIDDK NIH HHS, United States
    Id: R01-DK084391
  • Agency: NIH HHS, United States
    Id: P40 OD011102
  • Agency: NIH HHS, United States
    Id: U42 OD010921
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK084391

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This is a list of tools and resources that we have found mentioned in this publication.


Mutant Mouse Resource and Research Center (tool)

RRID:SCR_002953

National public repository system for mutant mice. Archives and distributes scientifically valuable spontaneous and induced mutant mouse strains and ES cell lines for use by biomedical research community. Includes breeding/distribution facilities and information coordinating center. Mice strains are cryopreserved, unless live colony must be established. Live mice are supplied from production colony, from colony recovered from cryopreservation, or via micro-injection of cell line into host blastocysts. MMRRC member facilities also develop technologies to improve handling of mutant mice, including advances in assisted reproductive techniques, cryobiology, genetic analysis, phenotyping and infectious disease diagnostics.

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Mutant Mouse Resource and Research Center - Jackson Laboratory (tool)

RRID:SCR_016446

Center for mutant mouse research and distribution. The objectives of the JAX MMRRC are to: identify and evaluate biomedically-significant mice, import/acquire and archive mouse strains, distribute mouse strains, and operate a control program to ensure genetic stability.

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