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JoVE Biology

Michael J. Boland1, Jennifer L. Hazen1, Kristopher L. Nazor1, Alberto R. Rodriguez2, Greg Martin2, Sergey Kupriyanov2, Kristin K. Baldwin1

1Dorris Neuroscience Center & Department of Cell Biology, The Scripps Research Institute, 2Mouse Genetics Core Facility, The Scripps Research Institute

Generating induced pluripotent stem cell (iPSC) lines produces lines of differing developmental potential even when they pass standard tests for pluripotency. Here we describe a protocol to produce mice derived entirely from iPSCs, which defines the iPSC lines as possessing full pluripotency1.

Date Published: 11/29/2012, Issue 69; doi: 10.3791/4003

Keywords: Stem Cell Biology, Issue 69, Molecular Biology, Developmental Biology, Medicine, Cellular Biology, Induced pluripotent stem cells, iPSC, stem cells, reprogramming, developmental potential, tetraploid embryo complementation, mouse

Boland, M. J., Hazen, J. L., Nazor, K. L., Rodriguez, A. R., Martin, G., Kupriyanov, S., et al. Generation of Mice Derived from Induced Pluripotent Stem Cells. J. Vis. Exp. (69), e4003, doi:10.3791/4003 (2012).

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also produce a source of patient-matched cells for regenerative therapies. iPSCs may be generated using multiple protocols and derived from multiple cell sources. Once generated, iPSCs are tested using a variety of assays including immunostaining for pluripotency markers, generation of three germ layers in embryoid bodies and teratomas, comparisons of gene expression with embryonic stem cells (ESCs) and production of chimeric mice with or without germline contribution2. Importantly, iPSC lines that pass these tests still vary in their capacity to produce different differentiated cell types2. This has made it difficult to establish which iPSC derivation protocols, donor cell sources or selection methods are most useful for different applications.

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Generating Mice from Induced Pluripotent Stem Cells | Protocol