Defined conditions for propagation and manipulation of mouse embryonic stem cells.
Development (Cambridge, England)
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Mulas, C., Kalkan, T., von Meyenn, F., Leitch, H. G., Nichols, J., & Smith, A. (2019). Defined conditions for propagation and manipulation of mouse embryonic stem cells.. Development (Cambridge, England), 146 (6)https://doi.org/10.1242/dev.173146
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 aberration and genetic abnormality. Here we provide methodology 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.
Neurons, Animals, Mice, Inbred C57BL, Humans, Mice, RNA, Small Interfering, Culture Media, Cell Culture Techniques, Coculture Techniques, Karyotyping, Signal Transduction, Cell Cycle, Cell Differentiation, Embryonic Stem Cells, Embryo, Mammalian, CRISPR-Cas Systems, Mouse Embryonic Stem Cells
The funding statement is uploaded separately from the manuscript but the authors acknowledged Wellcome Trust, BBSRC and MRC. Austin Smith is an MRC Professor.
Wellcome Trust (091484/Z/10/Z)
External DOI: https://doi.org/10.1242/dev.173146
This record's URL: https://www.repository.cam.ac.uk/handle/1810/289900