Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin.

Autor:	Zhang J; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States.; Stem Cell and Regenerative Medicine Center, University of Wisconsin - Madison, Madison, United States., Gregorich ZR; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States., Tao R; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States., Kim GC; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States., Lalit PA; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States., Carvalho JL; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States.; Department of Genomic Sciences and Biotechnology, University of Brasília, Brasília, Brazil., Markandeya Y; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States., Mosher DF; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States.; Morgridge Institute for Research, Madison, United States.; Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, United States., Palecek SP; Stem Cell and Regenerative Medicine Center, University of Wisconsin - Madison, Madison, United States.; Department of Chemical and Biological Engineering, College of Engineering, University of Wisconsin, Madison, United States., Kamp TJ; Department of Medicine, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States.; Stem Cell and Regenerative Medicine Center, University of Wisconsin - Madison, Madison, United States.; Department of Cell and Regenerative Biology, School of Medicine and Public Health, University of Wisconsin - Madison, Madison, United States.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2022 Jun 27; Vol. 11. Date of Electronic Publication: 2022 Jun 27.
DOI:	10.7554/eLife.69028
Abstrakt:	Research and therapeutic applications using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) require robust differentiation strategies. Efforts to improve hPSC-CM differentiation have largely overlooked the role of extracellular matrix (ECM). The present study investigates the ability of defined ECM proteins to promote hPSC cardiac differentiation. Fibronectin (FN), laminin-111, and laminin-521 enabled hPSCs to attach and expand. However, only addition of FN promoted cardiac differentiation in response to growth factors Activin A, BMP4, and bFGF in contrast to the inhibition produced by laminin-111 or laminin-521. hPSCs in culture produced endogenous FN which accumulated in the ECM to a critical level necessary for effective cardiac differentiation. Inducible shRNA knockdown of FN prevented Brachyury + mesoderm formation and subsequent hPSC-CM generation. Antibodies blocking FN binding integrins α4β1 or αVβ1, but not α5β1, inhibited cardiac differentiation. Furthermore, inhibition of integrin-linked kinase led to a decrease in phosphorylated AKT, which was associated with increased apoptosis and inhibition of cardiac differentiation. These results provide new insights into defined matrices for culture of hPSCs that enable production of FN-enriched ECM which is essential for mesoderm formation and efficient cardiac differentiation. Competing Interests: JZ, ZG, RT, GK, PL, JC, YM, DM, SP No competing interests declared, TK is a consultant for Fujifilm Cellular Dynamics Incorporated, a stem cell company (© 2022, Zhang et al.)
Databáze:	MEDLINE
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