TY - JOUR
T1 - Human pluripotent stem cell culture density modulates YAP signaling
AU - Hsiao, Cheston
AU - Lampe, Michael
AU - Nillasithanukroh, Songkhun
AU - Han, Wenqing
AU - Lian, Xiaojun
AU - Palecek, Sean P.
N1 - Funding Information:
We thank the WiCell Research Institute and BIONAnocomposite Tissue Engineering Scaffolds theme in the Wisconsin Institutes for Discovery for providing cell lines, reagents and equipment. The PAX6 monoclonal antibody developed by Atsushi Kawaka was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. This work was supported by National Institutes of Health Grant R01EB007534.
Publisher Copyright:
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2016/5/1
Y1 - 2016/5/1
N2 - Human pluripotent stem cell (hPSC) density is an important factor in self-renewal and differentiation fates; however, the mechanisms through which hPSCs sense cell density and process this information in making cell fate decisions remain to be fully understood. One particular pathway that may prove important in density-dependent signaling in hPSCs is the Hippo pathway, which is regulated by cell-cell contact and mechanosensing through the cytoskeleton and has been linked to the maintenance of stem cell pluripotency. To probe regulation of Hippo pathway activity in hPSCs, we assessed whether Hippo pathway transcriptional activator YAP was differentially modulated by cell density. At higher cell densities, YAP phosphorylation and localization to the cytoplasm increased, which led to decreased YAP-mediated transcriptional activity. Furthermore, total YAP protein levels diminished at high cell density due to the phosphorylation-targeted degradation of YAP. Inducible shRNA knockdown of YAP reduced expression of YAP target genes and pluripotency genes. Finally, the density-dependent increase of neuroepithelial cell differentiation was mitigated by shRNA knockdown of YAP. Our results suggest a pivotal role of YAP in cell density-mediated fate decisions in hPSCs.
AB - Human pluripotent stem cell (hPSC) density is an important factor in self-renewal and differentiation fates; however, the mechanisms through which hPSCs sense cell density and process this information in making cell fate decisions remain to be fully understood. One particular pathway that may prove important in density-dependent signaling in hPSCs is the Hippo pathway, which is regulated by cell-cell contact and mechanosensing through the cytoskeleton and has been linked to the maintenance of stem cell pluripotency. To probe regulation of Hippo pathway activity in hPSCs, we assessed whether Hippo pathway transcriptional activator YAP was differentially modulated by cell density. At higher cell densities, YAP phosphorylation and localization to the cytoplasm increased, which led to decreased YAP-mediated transcriptional activity. Furthermore, total YAP protein levels diminished at high cell density due to the phosphorylation-targeted degradation of YAP. Inducible shRNA knockdown of YAP reduced expression of YAP target genes and pluripotency genes. Finally, the density-dependent increase of neuroepithelial cell differentiation was mitigated by shRNA knockdown of YAP. Our results suggest a pivotal role of YAP in cell density-mediated fate decisions in hPSCs.
UR - http://www.scopus.com/inward/record.url?scp=84975780433&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84975780433&partnerID=8YFLogxK
U2 - 10.1002/biot.201500374
DO - 10.1002/biot.201500374
M3 - Article
C2 - 26766309
AN - SCOPUS:84975780433
SN - 1860-6768
VL - 11
SP - 662
EP - 675
JO - Biotechnology Journal
JF - Biotechnology Journal
IS - 5
ER -