Direct cell reprogramming is a stochastic process amenable to acceleration.
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.
Hanna
Jacob
J
The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA. Hanna@wi.mit.edu
Saha
Krishanu
K
Pando
Bernardo
B
van Zon
Jeroen
J
Lengner
Christopher J
CJ
Creyghton
Menno P
MP
van Oudenaarden
Alexander
A
Jaenisch
Rudolf
R
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
C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors