title: Effects of endothelial growth media on proepicardial cell  gene expression and morphogenesis in 3D collagen matrices.
abstract: Proepicardial cells (PE) contribute to embryonic coronary vessel and epicardial development. Cells from the PE region can differentiate into coronary vascular smooth muscle cells and fibroblasts in vitro, but the endothelial specification capability of these cells is controversial. We sought to examine the effects of endothelial cell growth media on gene expression and the morphogenic properties of proepicardial cells in three-dimensional (3D) matrices. A primary culture of avian PE cells was subjected to molecular characterization with selected endothelial specific markers. Morphogenic properties of PE cells were assessed by in vitro assays of coronary vasculogenesis and invasion, which utilized highly defined, serum free, three-dimensional matrix conditions. PE cells maintained mixed cell population properties in the culture based on morphogenic features, immunohistochemistry, and the gene expression data. When suspended in a 3D vasculogenesis in vitro assay, PE cells formed intracellular vacuoles and assembled into multicellular tubes. Further, ultrastructural analysis revealed the presence of pinocytic vacuoles, intercellular junctions, and endothelial specific Weibel Palade bodies. In the invasion assay, PE cells spontaneously invaded control matrices. This invasion was markedly enhanced by lysophosphatidic acid (94±9.6 vs. 285.6±54.9, p<0.05) and was completely blocked with synthetic broad-spectrum metalloproteinase inhibitor GM6001. Isolated PE cells grown in endothelial cell media represent mixed-cell population, characterized by both smooth muscle and endothelial gene expression. When placed in 3D in vitro assays, PE cells manifest morphogenic properties, including multicellular tube assembly and invasion.
authors.affiliation: Department of Internal Medicine, Division of Cardiovascular Medicine, University of Missouri School of Medicine, Columbia, MO, USA. karuparthip@health.missouri.edu
authors.forename: Poorna
authors.initials: P
authors.lastname: Karuparthi
authors.forename: Karen
authors.initials: K
authors.lastname: Nickelson
authors.forename: Dmitri
authors.initials: D
authors.lastname: Baklanov
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D002642:Chick Embryo; D003094:Collagen; D003470:Culture Media; D005109:Extracellular Matrix / Q000378:metabolism / Q000502:physiology; D015870:Gene Expression / Q000187:drug effects*; D046529:Microscopy, Electron, Transmission; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D010496:Pericardium / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D054457:Tissue Scaffolds
chemical_list: D003470:Culture Media; D003094:Collagen
references.citation: In Vitro Cell Dev Biol Anim. 2000 Sep;36(8):513-9
references.pmid: 11149750
references.citation: Microvasc Res. 2007 Mar;73(2):84-94
references.pmid: 17223139
references.citation: Proc Natl Acad Sci U S A. 1979 Nov;76(11):5674-8
references.pmid: 293671
references.citation: Dev Dyn. 2007 Mar;236(3):684-95
references.pmid: 17238175
references.citation: Endothelium. 2003;10(4-5):217-23
references.pmid: 14660081
references.citation: Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9504-8
references.pmid: 1409660
references.citation: Dev Dyn. 1992 Dec;195(4):231-72
references.pmid: 1304821
references.citation: Circulation. 2007 Aug 21;116(8):917-27
references.pmid: 17684151
references.citation: J Cell Biol. 2004 Mar 15;164(6):811-7
references.pmid: 15024030
references.citation: Dev Biol. 1989 Mar;132(1):230-40
references.pmid: 2783918
references.citation: Nature. 2007 Jan 11;445(7124):177-82
references.pmid: 17108969
references.citation: Nature. 2005 Sep 15;437(7057):426-31
references.pmid: 16163360
references.citation: J Cell Biol. 1984 Dec;99(6):2034-40
references.pmid: 6501412
references.citation: Dev Dyn. 2006 Jan;235(1):19-28
references.pmid: 16145662
references.citation: Exp Cell Res. 1996 Apr 10;224(1):39-51
references.pmid: 8612690
references.citation: Physiol Rev. 2004 Jul;84(3):869-901
references.pmid: 15269339
references.citation: Nature. 2006 Jul 27;442(7101):453-6
references.pmid: 16799567
references.citation: Circ Res. 2005 May 27;96(10):1039-41
references.pmid: 15879310
references.citation: Mol Cancer. 2006 Dec 08;5:69
references.pmid: 17156449
references.citation: Lab Invest. 1958 Mar-Apr;7(2):134-7
references.pmid: 13540204
references.citation: J Cell Biol. 1964 Oct;23:101-12
references.pmid: 14228505
references.citation: Circ Res. 1993 Sep;73(3):559-68
references.pmid: 8348697