title: Direct cell reprogramming is a stochastic process amenable to acceleration.
abstract: Direct reprogramming of somatic cells into induced pluripotent stem (iPS) cells can be achieved by overexpression of Oct4, Sox2, Klf4 and c-Myc transcription factors, but only a minority of donor somatic cells can be reprogrammed to pluripotency. Here we demonstrate that reprogramming by these transcription factors is a continuous stochastic process where almost all mouse donor cells eventually give rise to iPS cells on continued growth and transcription factor expression. Additional inhibition of the p53/p21 pathway or overexpression of Lin28 increased the cell division rate and resulted in an accelerated kinetics of iPS cell formation that was directly proportional to the increase in cell proliferation. In contrast, Nanog overexpression accelerated reprogramming in a predominantly cell-division-rate-independent manner. Quantitative analyses define distinct cell-division-rate-dependent and -independent modes for accelerating the stochastic course of reprogramming, and suggest that the number of cell divisions is a key parameter driving epigenetic reprogramming to pluripotency.
authors.affiliation: The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA. Hanna@wi.mit.edu
authors.forename: Jacob
authors.initials: J
authors.lastname: Hanna
authors.forename: Krishanu
authors.initials: K
authors.lastname: Saha
authors.forename: Bernardo
authors.initials: B
authors.lastname: Pando
authors.forename: Jeroen
authors.initials: J
authors.lastname: van Zon
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lengner
authors.forename: Menno P
authors.initials: MP
authors.lastname: Creyghton
authors.forename: Alexander
authors.initials: A
authors.lastname: van Oudenaarden
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D065150:Cellular Reprogramming; D018507:Gene Expression Regulation, Developmental; D000090062:Kruppel-Like Factor 4; D051379:Mice; D016513:Mice, SCID; D008954:Models, Biological; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors