title: Isolation of Reprogramming Intermediates During Generation of Induced Pluripotent Stem Cells from Mouse Embryonic Fibroblasts.
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).
authors.affiliation: Department of Anatomy and Developmental Biology, Australian Regenerative Medicine Institute, Monash University, Level 3, STRIP 1, Building 75, Wellington Road, Clayton, VIC, 3800, Australia.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Nefzger
authors.affiliation: Department of Anatomy and Developmental Biology, Australian Regenerative Medicine Institute, Monash University, Level 3, STRIP 1, Building 75, Wellington Road, Clayton, VIC, 3800, Australia.
authors.forename: Sara
authors.initials: S
authors.lastname: Alaei
authors.affiliation: Department of Anatomy and Developmental Biology, Australian Regenerative Medicine Institute, Monash University, Level 3, STRIP 1, Building 75, Wellington Road, Clayton, VIC, 3800, Australia. jose.polo@monash.edu.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Polo
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D065150:Cellular Reprogramming; D005347:Fibroblasts / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice
keywords: Cell surface marker; Fluorescence-activated cell sorting; Induced pluripotent stem cells; Reprogrammable mouse model; Reprogramming intermediates