title: Expansion and patterning of cardiovascular progenitors derived from human pluripotent stem cells.
abstract: The inability of multipotent cardiovascular progenitor cells (CPCs) to undergo multiple divisions in culture has precluded stable expansion of precursors of cardiomyocytes and vascular cells. This contrasts with neural progenitors, which can be expanded robustly and are a renewable source of their derivatives. Here we use human pluripotent stem cells bearing a cardiac lineage reporter to show that regulated MYC expression enables robust expansion of CPCs with insulin-like growth factor-1 (IGF-1) and a hedgehog pathway agonist. The CPCs can be patterned with morphogens, recreating features of heart field assignment, and controllably differentiated to relatively pure populations of pacemaker-like or ventricular-like cardiomyocytes. The cells are clonogenic and can be expanded for >40 population doublings while retaining the ability to differentiate into cardiomyocytes and vascular cells. Access to CPCs will allow precise recreation of elements of heart development in vitro and facilitate investigation of the molecular basis of cardiac fate determination. This technology is applicable for cardiac disease modeling, toxicology studies and tissue engineering.
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Birket
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Marcelo C
authors.identifier: http://orcid.org/0000-0003-0583-7443
authors.initials: MC
authors.lastname: Ribeiro
authors.affiliation: Amsterdam Medical Center, Amsterdam, the Netherlands.
authors.forename: Arie O
authors.initials: AO
authors.lastname: Verkerk
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Dorien
authors.initials: D
authors.lastname: Ward
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Ana Rita
authors.initials: AR
authors.lastname: Leitoguinho
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Sabine C
authors.initials: SC
authors.lastname: den Hartogh
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Valeria V
authors.initials: VV
authors.lastname: Orlova
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Harsha D
authors.identifier: http://orcid.org/0000-0001-5343-1021
authors.initials: HD
authors.lastname: Devalla
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Verena
authors.initials: V
authors.lastname: Schwach
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Milena
authors.initials: M
authors.lastname: Bellin
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Robert
authors.initials: R
authors.lastname: Passier
authors.affiliation: Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Christine L
authors.initials: CL
authors.lastname: Mummery
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D053823:Hedgehog Proteins