Defined conditions for propagation and manipulation of mouse embryonic stem cells

ABSTRACT The power of mouse embryonic stem (ES) cells to colonise the developing embryo has revolutionised mammalian developmental genetics and stem cell research. This power is vulnerable, however, to the cell culture environment, deficiencies in which can lead to cellular heterogeneity, adaptive phenotypes, epigenetic aberrations and genetic abnormalities. Here, we provide detailed methodologies for derivation, propagation, genetic modification and primary differentiation of ES cells in 2i or 2i+LIF media without serum or undefined serum substitutes. Implemented diligently, these procedures minimise variability and deviation, thereby improving the efficiency, reproducibility and biological validity of ES cell experimentation. Summary: Detailed protocols for the propagation, manipulation and differentiation of mouse embryonic stem cells, with tips for minimising variability and improving the efficiency, reproducibility and reliability of embryonic stem cell culture.

• Publication year

  2019

• Venue

  Development

• Publication date

  2019-03-15

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1242/dev.173146PMID 30914406PMCID 6451320
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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