Laminin-guided highly efficient endothelial commitment from human pluripotent stem cells.
Obtaining highly purified differentiated cells via directed differentiation from human pluripotent stem cells (hPSCs) is an essential step for their clinical application. Among the various conditions that should be optimized, the precise role and contribution of the extracellular matrix (ECM) during differentiation are relatively unclear. Here, using a short fragment of laminin 411 (LM411-E8), an ECM predominantly expressed in the vascular endothelial basement membrane, we demonstrate that the directed switching of defined ECMs robustly yields highly-purified (>95%) endothelial progenitor cells (PSC-EPCs) without cell sorting from hPSCs in an integrin-laminin axis-dependent manner. Single-cell RNA-seq analysis revealed that LM411-E8 resolved intercellular transcriptional heterogeneity and escorted the progenitor cells to the appropriate differentiation pathway. The PSC-EPCs gave rise to functional endothelial cells both in vivo and in vitro. We therefore propose that sequential switching of defined matrices is an important concept for guiding cells towards desired fate.
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Ryo
R
Ohta
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Akira
A
Niwa
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita, Japan.
Yukimasa
Y
Taniguchi
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Naoya M
NM
Suzuki
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita, Japan.
Junko
J
Toga
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita, Japan.
Emiko
E
Yagi
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Norikazu
N
Saiki
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Yoko
Y
Nishinaka-Arai
Depeartment of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.|Mitsubishi Space Software CO., LTD, Amagasaki, Japan.
Chihiro
C
Okada
Depeartment of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Akira
A
Watanabe
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Tatsutoshi
T
Nakahata
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita, Japan.
Kiyotoshi
K
Sekiguchi
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
Megumu K
MK
Saito
D001483:Base Sequence; D001485:Basement Membrane / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology*; D061252:Feeder Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D039121:Integrin alpha6beta1 / Q000378:metabolism; D007797:Laminin / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D017423:Sequence Analysis, RNA
D039121:Integrin alpha6beta1; D007797:Laminin; D010446:Peptide Fragments