title: Isolation of endothelial cells and pericytes from swine corpus luteum.
abstract: From an angiogenesis perspective, the ovary offers a unique opportunity to study the physiological development of blood vessels. The first purpose of this work was to set up a protocol for the isolation of pig corpus luteum endothelial cells, which were characterized by both morphologic parameters and the expression of typical molecular markers; we also verified their ability to form capillary-like structures in a 3-dimensional matrix, their response to hypoxia and their migration in the presence of vascular endothelial growth factor (VEGF). The effectiveness of our isolation protocol was confirmed by the characteristic "cobblestone shape" of isolated cells at confluence as well as their expression of all the examined endothelial markers. Our data also showed a significant cell production of VEGF and nitric oxide. Isolated endothelial cells were also responsive to hypoxia by increasing the expression and production of VEGF and decreasing that of nitric oxide. In the angiogenesis bioassay, cells displayed the ability of forming capillary-like structures and also exhibited a significant migration in the scratch test. Our data suggest that the isolation of luteal endothelial cells represents a promising tool in experiments designed to clarify the biology of the angiogenic process. Furthermore, we have demonstrated that the isolated population comprises a subset of cells with a multidifferentiative capacity toward the chondrocytic and adipocytic phenotypes. These data suggest the presence of a perivascular or adventitial cell niche in the vascular wall of the corpus luteum populated with cells showing mesenchymal stem cell-like features, as already demonstrated for the adipose tissue and endometrium.
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy. Electronic address: giuseppina.basini@unipr.it.
authors.forename: G
authors.initials: G
authors.lastname: Basini
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Falasconi
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Bussolati
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Grolli
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Ramoni
authors.affiliation: Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Grasselli
mesh_terms: D050156:Adipogenesis; D000818:Animals; D002454:Cell Differentiation; D002465:Cell Movement / Q000502:physiology; D020219:Chondrogenesis; D003338:Corpus Luteum / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology*; D005260:Female; D005786:Gene Expression Regulation; D018919:Neovascularization, Physiologic; D010012:Osteogenesis; D010100:Oxygen; D020286:Pericytes / Q000166:cytology*; D013552:Swine / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: D042461:Vascular Endothelial Growth Factor A; D010100:Oxygen
keywords: Angiogenesis; NO; Ovary; Pericytes; VEGF