Export
Transforming growth factor beta 1 modulates extracellular matrix organization and cell-cell junctional complex formation during in vitro angiogenesis
Periodical: J Cell Physiol ISBN: 0021-9541 (Print)
Number: 1
Date: 1990/01/01
Language: eng
Pages: 117-28
Authors:Merwin, J. R., Anderson, J. M., Kocher, O., Van Itallie, C. M., Madri, J. A.
A copy of this paper may be available for free: Google Scholar Search 
Abstract
Transforming growth factor-beta 1 (TGF-beta 1) is angiogenic in vivo. In two-dimensional (2-D) culture systems microvascular endothelial cell proliferation is inhibited up to 80% by TGF-beta 1; however, in three-dimensional (3-D) collagen gels TGF-beta 1 is found to have no effect on proliferation while eliciting the formation of calcium and magnesium dependent tube-like structures mimicking angiogenesis. DNA analyses performed on 3-D cell cultures reveal no significant difference in the amount of DNA or cell number in control versus TGF-beta 1 treated cultures. In 2-D cultures TGF-beta 1 is known to increase cellular fibronectin accumulation; however, in 3-D cultures no difference is seen between control and TGF-beta 1 treated cells as established by ELISA testing for type IV collagen, fibronectin, and laminin. In 3-D cultures there is increased synthesis and secretion of type V collagen in both control and TGF-beta 1 treated cultures over 2-D cultures. Even though an equal amount of type V collagen is seen in both 3-D conditions, there is a reorganization of the protein with concentration along an organizing basal lamina in TGF-beta 1 treated cultures. EM morphological analyses on 3-D cultures illustrate quiescent, control cells lacking cell contacts. In contrast, TGF-beta 1 treated cells show increased pseudopod formation, cell-cell contact, and organized basal lamina-like material closely apposed to the "abluminal" plasma membranes. TGF-beta 1 treated cells also appear to form junctional complexes between adjoining cells. Immunofluorescence using specific antibodies to the tight junction protein ZO-1 results in staining at apparent cell-cell junctions in the 3-D cultures. Northern blots of freshly isolated microvascular endothelium, 2-D and 3-D cultures, using cDNA and cRNA probes specific for the ZO-1 tight junction protein, reveal the presence of the 7.8 kb mRNA. Western blots of rat epididymal fat pad endothelial cells (RFC) monolayer lysates probed with anti
Transforming growth factor-beta 1 (TGF-beta 1) is angiogenic in vivo. In two-dimensional (2-D) culture systems microvascular endothelial cell proliferation is inhibited up to 80% by TGF-beta 1; however, in three-dimensional (3-D) collagen gels TGF-beta 1 is found to have no effect on proliferation while eliciting the formation of calcium and magnesium dependent tube-like structures mimicking angiogenesis. DNA analyses performed on 3-D cell cultures reveal no significant difference in the amount of DNA or cell number in control versus TGF-beta 1 treated cultures. In 2-D cultures TGF-beta 1 is known to increase cellular fibronectin accumulation; however, in 3-D cultures no difference is seen between control and TGF-beta 1 treated cells as established by ELISA testing for type IV collagen, fibronectin, and laminin. In 3-D cultures there is increased synthesis and secretion of type V collagen in both control and TGF-beta 1 treated cultures over 2-D cultures. Even though an equal amount of type V collagen is seen in both 3-D conditions, there is a reorganization of the protein with concentration along an organizing basal lamina in TGF-beta 1 treated cultures. EM morphological analyses on 3-D cultures illustrate quiescent, control cells lacking cell contacts. In contrast, TGF-beta 1 treated cells show increased pseudopod formation, cell-cell contact, and organized basal lamina-like material closely apposed to the "abluminal" plasma membranes. TGF-beta 1 treated cells also appear to form junctional complexes between adjoining cells. Immunofluorescence using specific antibodies to the tight junction protein ZO-1 results in staining at apparent cell-cell junctions in the 3-D cultures. Northern blots of freshly isolated microvascular endothelium, 2-D and 3-D cultures, using cDNA and cRNA probes specific for the ZO-1 tight junction protein, reveal the presence of the 7.8 kb mRNA. Western blots of rat epididymal fat pad endothelial cells (RFC) monolayer lysates probed with anti
Keywords
Cell Culture Techniques/*methods, Cell Division, Cell Line, Coculture Techniques, Collagen Type IV/metabolism, Cyclin-Dependent Kinase Inhibitor p21, Cyclins/metabolism, Extracellular Matrix, Fibroblasts/cytology/metabolism, Humans, Keratinocytes/*cytology/*metabolism, Keratins/metabolism, Mesoderm/cytology/metabolism, Phenotype, Proliferating Cell Nuclear Antigen/metabolism, Tumor Suppressor Protein p53/metabolism
Cell Culture Techniques/*methods, Cell Division, Cell Line, Coculture Techniques, Collagen Type IV/metabolism, Cyclin-Dependent Kinase Inhibitor p21, Cyclins/metabolism, Extracellular Matrix, Fibroblasts/cytology/metabolism, Humans, Keratinocytes/*cytology/*metabolism, Keratins/metabolism, Mesoderm/cytology/metabolism, Phenotype, Proliferating Cell Nuclear Antigen/metabolism, Tumor Suppressor Protein p53/metabolism
3DCellculture.com's MAMI
search attributes
CellLine: Primary-hSkinEC
Morphology: Endothelial
Origin: Skin
Species: Human
Scaffold Form: gel/hydrogelMorphology: Endothelial
Origin: Skin
Species: Human
Scaffold Material: Collagen