Derivation and maintenance of mouse haploid embryonic stem cells.
Forment, Josep V
Vicente, Jose R
Adams, David J
Nature Publishing Group
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Elling, U., Woods, M., Forment, J. V., Fu, B., Yang, F., Ng, B. L., Vicente, J. R., et al. (2019). Derivation and maintenance of mouse haploid embryonic stem cells.. Nature protocols, 14 (7), 1991-2014. https://doi.org/10.1038/s41596-019-0169-z
Ploidy represents the number of chromosome sets in a cell. Whereas gametes have a haploid genome (n), most mammalian cells have diploid genomes (2n). The diploid status of most cells correlates to the number of probable alleles for each autosomal gene and makes it difficult to target these genes via mutagenesis techniques. Here we describe a seven-week protocol for the derivation of mouse haploid embryonic stem cells (hESC) from female gametes that also describes how to maintain the cells once derived. We also detail additional procedures that can be used with cell lines obtained from the mouse Haplobank, a biobank of over 100,000 individual mouse hESC lines with targeted mutations in 16,970 genes. hESC can spontaneously diploidize and can be maintained both in haploid and diploid states. Mouse hESC are genomically and karyotypically stable, are innately immortal, isogenic and can be derived in an array of differentiated cell types, and are thus highly amenable for genetic screens and for defining molecular connectivity pathways.
Cell Line, Blastocyst, Animals, Mice, Inbred Strains, Mice, Inbred C57BL, Mice, Cell Culture Techniques, Cell Separation, Haploidy, Female, Workflow, Mouse Embryonic Stem Cells
UK Dementia Research Institute fellowship (MC_PC_17111).
Cancer Research UK (A18796)
Wellcome Trust (206388/Z/17/Z)
Wellcome Trust (092096/Z/10/Z)
Cancer Research UK (A14492)
External DOI: https://doi.org/10.1038/s41596-019-0169-z
This record's URL: https://www.repository.cam.ac.uk/handle/1810/290997
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