PT  - JOURNAL ARTICLE
AU  - Cristina Esteva-Font
AU  - Tao Su
AU  - Caitlin Peaslee
AU  - Caroline Duwaerts
AU  - Ke Liu
AU  - Marisa Medina
AU  - Jacquelyn J. Maher
AU  - Aras N. Mattis
TI  - Doxycycline Significantly Enhances Induction of iPSCs to Endoderm by Enhancing survival via AKT Phosphorylation
AID  - 10.1101/2020.04.13.034595
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.04.13.034595
4099  - http://biorxiv.org/content/early/2020/04/14/2020.04.13.034595.short
4100  - http://biorxiv.org/content/early/2020/04/14/2020.04.13.034595.full
AB  - Induced pluripotent stem cells (iPSCs) provide an important tool for the generation patient-derived cells including hepatocyte-like cells via developmental cues through an endoderm intermediate. However, most iPSCs fail to differentiate into endoderm, with induction resulting in apoptosis. To address this issue, we built upon published methods to develop an improved protocol with our discovery that doxycycline dramatically enhances the iPSC to endoderm differentiation efficiency by inhibiting apoptosis and promoting proliferation via the AKT pathway. We tested this new protocol in more than 70 iPSC lines with consistent formation of complete sheets of endoderm in 90%. Endoderm generated by our method achieves similar transcriptomic profiles, including FOXA2, HNF1β, CXCR4, and SOX17 positive cells, and the ability to be further differentiated. Furthermore this method achieves a four-fold increase in endoderm cell number and will accelerate studies of human diseases in vitro and facilitate the expansion of iPSC-derived cells for transplantation studies.Competing Interest StatementA.N.M. is a consultant for HepaTx and Ambys Medicines. The University of California San Francisco, has filed U.S. Provisional Patent Application No. 62/947,308.