Defined conditions for propagation and manipulation of mouse embryonic stem cells

Carla Mulas; Tüzer Kalkan; Ferdinand von Meyenn; Harry G. Leitch; Jennifer Nichols; Austin Smith

doi:10.1242/dev.173146

Defined conditions for propagation and manipulation of mouse embryonic stem cells

Carla Mulas, Tüzer Kalkan, Ferdinand von Meyenn, Harry G. Leitch, Jennifer Nichols, Austin Smith

Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Gleeson Building, Tennis Court Road, Cambridge CB2 1QR, UK.
King's College London
Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London W12 0NN, UK.
Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3DY, UK.
Department of Biochemistry, University of Cambridge, Hopkins Building, Tennis Court Road, Cambridge CB2 1QW, UK.

Research output: Contribution to journal › Article › peer-review

67 Citations (Scopus)

283 Downloads (Pure)

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.

Original language	English
Journal	Development
Volume	146
Issue number	6
Early online date	26 Mar 2019
DOIs	https://doi.org/10.1242/dev.173146
Publication status	Published - 26 Mar 2019

Keywords

Differentiation
Embryonic stem cells
Pluripotency
Self-renewal

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Defined conditions for propagation_MULAS_Accepted19February2019_GOLD VoR (CC BY)Final published version, 4.32 MBLicence: CC BY

Cite this

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AU - Smith, Austin

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Defined conditions for propagation and manipulation of mouse embryonic stem cells

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Keywords

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