Defined conditions for propagation and manipulation of mouse embryonic stem cells

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...

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Published inDevelopment (Cambridge) Vol. 146; no. 6
Main Authors Mulas, Carla, Kalkan, Tüzer, von Meyenn, Ferdinand, Leitch, Harry G., Nichols, Jennifer, Smith, Austin
Format Journal Article
LanguageEnglish
Published England The Company of Biologists Ltd 26.03.2019
Subjects
Animals
Cell Culture Techniques
Cell Cycle
Cell Differentiation - genetics
Coculture Techniques
CRISPR-Cas Systems
Culture Media - chemistry
Embryo, Mammalian - cytology
Embryonic Stem Cells - cytology
Humans
Karyotyping
Mice
Mice, Inbred C57BL
Mouse Embryonic Stem Cells - cytology
Neurons - cytology
RNA, Small Interfering - genetics
Signal Transduction
Stem Cells and Regeneration
Self-renewal
Differentiation
Embryonic stem cells
Pluripotency
Online AccessGet full text
ISSN0950-1991
1477-9129
1477-9129
DOI10.1242/dev.173146

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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.
AbstractList 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.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.
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.
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.
Author Leitch, Harry G.
Kalkan, Tüzer
Nichols, Jennifer
Smith, Austin
von Meyenn, Ferdinand
Mulas, Carla
AuthorAffiliation 6 Department of Biochemistry , University of Cambridge , Hopkins Building, Tennis Court Road, Cambridge CB2 1QW , UK
3 MRC London Institute of Medical Sciences (LMS) , Du Cane Road, London W12 0NN , UK
1 Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge , Gleeson Building, Tennis Court Road, Cambridge CB2 1QR , UK
2 Department of Medical and Molecular Genetics , King's College London , London SE1 9RT , UK
4 Institute of Clinical Sciences (ICS) , Faculty of Medicine , Imperial College London , Du Cane Road, London W12 0NN , UK
5 Department of Physiology, Development and Neuroscience , University of Cambridge , Downing Street, Cambridge CB2 3DY , UK
AuthorAffiliation_xml – name: 1 Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge , Gleeson Building, Tennis Court Road, Cambridge CB2 1QR , UK
– name: 2 Department of Medical and Molecular Genetics , King's College London , London SE1 9RT , UK
– name: 6 Department of Biochemistry , University of Cambridge , Hopkins Building, Tennis Court Road, Cambridge CB2 1QW , UK
– name: 3 MRC London Institute of Medical Sciences (LMS) , Du Cane Road, London W12 0NN , UK
– name: 5 Department of Physiology, Development and Neuroscience , University of Cambridge , Downing Street, Cambridge CB2 3DY , UK
– name: 4 Institute of Clinical Sciences (ICS) , Faculty of Medicine , Imperial College London , Du Cane Road, London W12 0NN , UK
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/30914406$$D View this record in MEDLINE/PubMed
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Issue 6
Keywords Self-renewal
Differentiation
Embryonic stem cells
Pluripotency
Language English
License http://creativecommons.org/licenses/by/4.0
2019. Published by The Company of Biologists Ltd.
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Snippet The power of mouse embryonic stem (ES) cells to colonise the developing embryo has revolutionised mammalian developmental genetics and stem cell research. This...
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SubjectTerms Animals
Cell Culture Techniques
Cell Cycle
Cell Differentiation - genetics
Coculture Techniques
CRISPR-Cas Systems
Culture Media - chemistry
Embryo, Mammalian - cytology
Embryonic Stem Cells - cytology
Humans
Karyotyping
Mice
Mice, Inbred C57BL
Mouse Embryonic Stem Cells - cytology
Neurons - cytology
RNA, Small Interfering - genetics
Signal Transduction
Stem Cells and Regeneration
Title Defined conditions for propagation and manipulation of mouse embryonic stem cells
URI https://www.ncbi.nlm.nih.gov/pubmed/30914406
https://www.proquest.com/docview/2198559015
https://pubmed.ncbi.nlm.nih.gov/PMC6451320
Volume 146
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