TY - JOUR
T1 - Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations
AU - Loh, Kyle M
AU - Ang, Lay Teng
AU - Zhang, Jingyao
AU - Kumar, Vibhor
AU - Ang, Jasmine
AU - Auyeong, Jun Qiang
AU - Lee, Kian Leong
AU - Choo, Siew Hua
AU - Lim, Christina Y Y
AU - Nichane, Massimo
AU - Tan, Junru
AU - Noghabi, Monireh Soroush
AU - Azzola, Lisa
AU - Ng, Elizabeth Siew-Sun
AU - Duruthy-Duruthy, Jens
AU - Sebastiano, Vittorio
AU - Poellinger, Lorenz
AU - Elefanty, Andrew G
AU - Stanley, Edouard
AU - Qingfeng, Chen
AU - Prabhakar, Shyam
AU - Weissman, Irving L
AU - Lim, Bing
PY - 2014
Y1 - 2014
N2 - Human pluripotent stem cell (hPSC) differentiation typically yields heterogeneous populations. Knowledge of signals controlling embryonic lineage bifurcations could efficiently yield desired cell types through exclusion of alternate fates. Therefore, we revisited signals driving induction and anterior-posterior patterning of definitive endoderm to generate a coherent roadmap for endoderm differentiation. With striking temporal dynamics, BMP and Wnt initially specified anterior primitive streak (progenitor to endoderm), yet, 24 hr later, suppressed endoderm and induced mesoderm. At lineage bifurcations, cross-repressive signals separated mutually exclusive fates; TGF-beta and BMP/MAPK respectively induced pancreas versus liver from endoderm by suppressing the alternate lineage. We systematically blockaded alternate fates throughout multiple consecutive bifurcations, thereby efficiently differentiating multiple hPSC lines exclusively into endoderm and its derivatives. Comprehensive transcriptional and chromatin mapping of highly pure endodermal populations revealed that endodermal enhancers existed in a surprising diversity of pre-enhancer states before activation, reflecting the establishment of a permissive chromatin landscape as a prelude to differentiation.
AB - Human pluripotent stem cell (hPSC) differentiation typically yields heterogeneous populations. Knowledge of signals controlling embryonic lineage bifurcations could efficiently yield desired cell types through exclusion of alternate fates. Therefore, we revisited signals driving induction and anterior-posterior patterning of definitive endoderm to generate a coherent roadmap for endoderm differentiation. With striking temporal dynamics, BMP and Wnt initially specified anterior primitive streak (progenitor to endoderm), yet, 24 hr later, suppressed endoderm and induced mesoderm. At lineage bifurcations, cross-repressive signals separated mutually exclusive fates; TGF-beta and BMP/MAPK respectively induced pancreas versus liver from endoderm by suppressing the alternate lineage. We systematically blockaded alternate fates throughout multiple consecutive bifurcations, thereby efficiently differentiating multiple hPSC lines exclusively into endoderm and its derivatives. Comprehensive transcriptional and chromatin mapping of highly pure endodermal populations revealed that endodermal enhancers existed in a surprising diversity of pre-enhancer states before activation, reflecting the establishment of a permissive chromatin landscape as a prelude to differentiation.
UR - http://www.sciencedirect.com/science/article/pii/S1934590913005560
U2 - 10.1016/j.stem.2013.12.007
DO - 10.1016/j.stem.2013.12.007
M3 - Article
SN - 1934-5909
VL - 14
SP - 237
EP - 252
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 2
ER -