Isolation of reprogramming intermediates during generation of induced pluripotent stem cells from mouse embryonic fibroblasts

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Abstract

Mature cells of the body can be reprogrammed towards a pluripotent state by forced expression of the transcription factors Oct-4, Klf-4, Sox2, and C-Myc (OKSM) at very low efficiency. To study the reprogramming process in detail the rare intermediates of the reaction need to be separated from the bulk population. Using a genetically engineered reprogrammable mouse strain we describe how to isolate intermediates from reprogramming cultures of mouse embryonic fibroblasts via antibody labeling of cell surface markers and fluorescence-activated cell sorting (FACS).
Original languageEnglish
Title of host publicationCell Reprogramming
Subtitle of host publicationMethods and Protocols
EditorsPaul J Verma, Huseyin Sumer
Place of PublicationTotowa NJ USA
PublisherHumana Press
Pages205 - 218
Number of pages14
Edition1st
ISBN (Print)9781493928477
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology: Springer Protocols
PublisherSpringer
Volume1330
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Cite this

Nefzger, C. M., Alaei Shehni, S., & Polo, J. M. (2015). Isolation of reprogramming intermediates during generation of induced pluripotent stem cells from mouse embryonic fibroblasts. In P. J. Verma, & H. Sumer (Eds.), Cell Reprogramming: Methods and Protocols (1st ed., pp. 205 - 218). (Methods in Molecular Biology: Springer Protocols; Vol. 1330). Humana Press. https://doi.org/10.1007/978-1-4939-2848-4_17