Transient p53 suppression increases reprogramming of human fibroblasts without affecting apoptosis and DNA damage

TY - JOUR

T1 - Transient p53 suppression increases reprogramming of human fibroblasts without affecting apoptosis and DNA damage

AU - Rasmussen, Mikkel Aabech

AU - Holst, Bjørn

AU - Tümer, Zeynep

AU - Johnsen, Mads G.

AU - Zhou, Shuling

AU - Stummann, Tina C.

AU - Hyttel, Poul

AU - Clausen, Christian

N1 - Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

PY - 2014/9/9

Y1 - 2014/9/9

N2 - The discovery of human-induced pluripotent stem cells (iPSCs) has sparked great interest in the potential treatment of patients with their own in vitro differentiated cells. Recently, knockout of the Tumor Protein 53 (p53) gene was reported to facilitate reprogramming but unfortunately also led to genomic instability. Here, we report that transient suppression of p53 during nonintegrative reprogramming of human fibroblasts leads to a significant increase in expression of pluripotency markers and overall number of iPSC colonies, due to downstream suppression of p21, without affecting apoptosis and DNA damage. Stable iPSC lines generated with or without p53 suppression showed comparable expression of pluripotency markers and methylation patterns, displayed normal karyotypes, contained between 0 and 5 genomic copy number variations and produced functional neurons in vitro. In conclusion, transient p53 suppression increases reprogramming efficiency without affecting genomic stability, rendering the method suitable for in vitro mechanistic studies with the possibility for future clinical translation.

AB - The discovery of human-induced pluripotent stem cells (iPSCs) has sparked great interest in the potential treatment of patients with their own in vitro differentiated cells. Recently, knockout of the Tumor Protein 53 (p53) gene was reported to facilitate reprogramming but unfortunately also led to genomic instability. Here, we report that transient suppression of p53 during nonintegrative reprogramming of human fibroblasts leads to a significant increase in expression of pluripotency markers and overall number of iPSC colonies, due to downstream suppression of p21, without affecting apoptosis and DNA damage. Stable iPSC lines generated with or without p53 suppression showed comparable expression of pluripotency markers and methylation patterns, displayed normal karyotypes, contained between 0 and 5 genomic copy number variations and produced functional neurons in vitro. In conclusion, transient p53 suppression increases reprogramming efficiency without affecting genomic stability, rendering the method suitable for in vitro mechanistic studies with the possibility for future clinical translation.

U2 - 10.1016/j.stemcr.2014.07.006

DO - 10.1016/j.stemcr.2014.07.006

M3 - Journal article

C2 - 25241739

SN - 2213-6711

VL - 3

SP - 404

EP - 413

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 3

ER -