|
Status |
Public on Feb 03, 2016 |
Title |
Derivation of Human Naïve Pluripotent Stem Cells by Activation of YAP |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by array
|
Summary |
Expression profiling of Naïve and Primed hESCs using Illumina HT-12 v.4 expression BeadChip We report that over-expression of the transcription factor YAP in male and female human ESCs and iPSCs promotes the generation of naïve PSCs. YAP-induced naïve PSCs have a naïve-specific dome-like colony morphology, rapid clonal growth rate for more than 70 passages, normal karyotype, expression of pluripotency markers and ability to form teratomas. Lysophosphatidic acid (LPA) can substitute for YAP to generated transgene-free human naïve PSCs. We performed microarray analyses on hESCs and hIPSCs in primed and naïve ESC media and show that YAP overexpressing cells in naïve media (Ying et al 2008, Ludwig et al 2006, Theunnissen 2014) display a distinct naïve-like transcriptional profile and other hallmarks of the naïve state. These results uncover an unexpected role for YAP in the transition to the human naïve state, and provide a platform in which to further dissect this transition and probe early human embryology.
|
|
|
Overall design |
Six primed and six naïve human ESCs (hESCs) were profiled in microarray experiments. Primed samples consist of H1 and H9 hESCs and IMR-90 iPSCs with or without overexpression of YAP where indicated, in standard primed hESC conditions. Naïve samples consist of YAP overexpressing cell lines that were profiled after conversion to the naïve state (mTeSR or N2B27 medium where indicated). Two cell lines, H9 ESCs, and WIBR ESCs (Theunissen et al 2014) were also generated in mTeSR naïve conditions without YAP overexpression, when using the YAP activator, Lysophosphatidic acid (LPA) - (“H9 mTeSR+2iFL+LPA naïve” and “WIBR3 mTeSR+2iFL+LPA naïve”). RNA samples were taken from the described cell lines and used in microarray experiments.
|
|
|
Contributor(s) |
Percharde M, Qin H, Ramalho-Santos M |
Citation(s) |
26947063 |
|
Submission date |
May 25, 2015 |
Last update date |
Aug 13, 2018 |
Contact name |
Michelle Percharde |
E-mail(s) |
m.percharde@lms.mrc.ac.uk
|
Organization name |
MRC Laboratory of Medical Sciences
|
Department |
Imperial College London
|
Street address |
Du Cane Road
|
City |
London |
ZIP/Postal code |
W12 0NN |
Country |
United Kingdom |
|
|
Platforms (1) |
GPL10558 |
Illumina HumanHT-12 V4.0 expression beadchip |
|
Samples (12)
|
|
Relations |
BioProject |
PRJNA284845 |