In vitro studies on the expansion of endothelial cell monolayers on components of the basement membrane

Kirkpatrick, Charles James; Kampe, M; H, Rixen; Fischer, Eike G.; Ruchatz, D; Mittermayer, Ch.

doi:10.1007/bf02890073

Cited by 14 publications

(8 citation statements)

References 27 publications

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“…In addition, induction of the α5β1 integrin has been described on tumor-associated angiogenic blood vessels in mouse and chicken, and functional blockade of this integrin halts angiogenesis and tumor growth (Kim et al, 2000a, b). Extra support comes from in vitro studies showing that fibronectin is a strong promoter of endothelial cell survival, proliferation and migration, aspects of cell behavior intrinsic to angiogenesis (Ingber and Folkman, 1988;McIntosh et al, 1988;Kirkpatrick et al, 1990;Ingber, 1992). Recently we showed that fibronectin is extremely potent at promoting brain endothelial cell survival and proliferation, and that these effects are mediated via the α5β1 and αvβ3 integrins (Wang and Milner, 2006).…”

Section: Hypoxic Induction Of the α5β1 Integrin On Cerebral Capillariesmentioning

confidence: 99%

Increased expression of fibronectin and the α5β1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia

Milner

Hung

Erokwu

et al. 2008

Molecular and Cellular Neuroscience

120

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The extracellular matrix (ECM) is an important regulator of angiogenesis and vascular remodeling. We showed previously that angiogenic capillaries in the developing CNS express high levels of fibronectin and its receptor α5β1 integrin, and that this expression is developmentally downregulated. As cerebral hypoxia leads to an angiogenic response, we sought to determine whether angiogenic vessels in the adult CNS re-express fibronectin and the α5β1 integrin. Ten-week old mice were subject to hypobaric hypoxia for 0, 4, 7 and 14 days, and fibronectin/integrin expression examined. Fibronectin and the α5 integrin subunit were strongly upregulated on capillaries in the hypoxic CNS, with the effect maximal at the earliest time point examined (4 days). Immunofluorescent studies demonstrated that the α5 integrin was expressed by angiogenic endothelial cells. In light of the defined angiogenic role for fibronectin in other systems, this work suggests that induction of fibronectin-α5β1 integrin expression may be an important molecular switch driving angiogenesis in the hypoxic CNS.

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Section: Hypoxic Induction Of the α5β1 Integrin On Cerebral Capillariesmentioning

confidence: 99%

Increased expression of fibronectin and the α5β1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia

Milner

Hung

Erokwu

et al. 2008

Molecular and Cellular Neuroscience

120

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“…However, the experiments with HUVEC presented in this paper classify collagen type III as a 'high' promoter of EC expansion. In addition, Lm, which elicited high expansion rates of HUVEC, gave the lowest monolayer expansion with BCEC, the mean value being significantly lower than the control value [8], This difference is highlighted by expressing the expansion rate relative to the corresponding control value (uncoated substratum = 100%). Thus, for BCEC, Lm gave an expansion rate of 25%, compared with 142% for HUVEC.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, we have shown that the highest rate of monolayer expansion of BCEC was on collagen type IV-F, followed by Fn [8]. Fn would thus appear to be an excellent substratum for promoting the monolayer ex pansion of both HUVEC and BCEC.…”

Section: Discussionmentioning

confidence: 99%

“…In a previous publication we discussed the cell biolog ical mechanisms operative in the described expansion assay [8]. One of the important mechanisms is prolifera tion of the EC.…”

Section: Discussionmentioning

confidence: 99%

“…In a recent investigation we developed an in vitro assay to simulate the in vivo situation in which an endothelial defect is repaired by adjacent intact endothelium [8]. This assay enables the ability of an intact EC monolayer to expand over individual components of the BM or extra cellular matrix to be tested.…”

Section: Introductionmentioning

confidence: 99%

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Differential Expansion of Human Endothelial Monolayers on Basement Membrane and Interstitial Collagens, Laminin and Fibronectin in vitro

Kirkpatrick¹,

Kampe²,

Fischer³

et al. 1990

Pathobiology

Self Cite

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In this study the ability of a human endothelial cell monolayer to expand over specific components of the basement membrane and extracellular matrix was investigated over a 5-day period. The method was intended as a model to study the mechanisms of endothelial regeneration. All components were coated onto sterile coverslips at a concentration of 10 micrograms/ml. The highest expansion was obtained on fibronectin, laminin and collagen type III, all three being statistically significantly greater than on the uncoated control surface (0.002 greater than p greater than 0.0001). Collagens types I and IV and a high molecular weight fragment mixture of type IV (IV-F, consisting of 75, 120 and 140 kD fragments) elicited approximately similar expansion rates, significantly higher than the control (0.02 greater than p greater than 0.003), although significantly lower (approximately 15%) than collagen type III, fibronectin and laminin (p less than 0.001). The high monolayer expansion on collagen type III is surprising, as it is a relatively minor biosynthetic product of the endothelial cell. It could, however, be of significance in wound healing, in which endothelial cells come into contact with this interstitial collagen. In addition, the similar results obtained with collagens IV and IV-F indicate that expansion of the endothelial monolayer is not dependent on the integrity of the tetrameric structure of type-IV collagen.

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Interaction of human malignant melanoma tumor spheroids with endothelium and reconstituted basement membrane: Modulation by RGDS

Offner

Feichtinger

Bigalke³

et al. 1993

Intl Journal of Cancer

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Tumor-cell extravasation involves sequential adhesive interactions with vascular endothelium and the subendothelial basement membrane. We have established a 3-dimensional model in vitro to simulate these events and to elucidate targets of the anti-cell-adhesive synthetic peptide RGDS. Tumor spheroids of the melanoma cell line ST-ML-12 served as models of tumor-cell emboli and were transferred onto human umbilical vein endothelial cells. To imitate the vascular anatomy, the latter were grown on reconstituted basement membranes produced by dextran-stimulated bovine corneal endothelial cells. RGDS did not affect the homotypic aggregation of the tumor cells and only minimally inhibited the attachment of the spheroids to the reconstituted vessel. A short-term (20 min) inhibition of adhesion to denuded basement membranes was observed. The attachment was closely associated with damage to the endothelial cells by oxygen-derived free radicals. RGDS retarded endothelial injury for up to 3 hr. The most prominent effect was observed after penetration of the endothelium. RGDS suppressed the emigration of tumor cells from the attached tumor-cell cluster in a dose- and time-dependent fashion. After 12 hr, the inhibitory effect progressively declined. This was not due to loss of activity of the peptide, indicating a resistance mechanism in the melanoma cells. On purified components of the basement membrane, RGDS effectively inhibited the initial spheroid attachment to fibronectin and collagen IV but had no effect on attachment to laminin. By contrast, subsequent migration was significantly suppressed on all substrata. Our model permits the study of dynamic cell-cell and cell-extracellular-matrix interactions and indicates that RGDS might predominantly act on early tumor-cell locomotion after penetration of the endothelium.

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In vitro studies on the expansion of endothelial cell monolayers on components of the basement membrane

Cited by 14 publications

References 27 publications

Increased expression of fibronectin and the α5β1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia

Increased expression of fibronectin and the α5β1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia

Differential Expansion of Human Endothelial Monolayers on Basement Membrane and Interstitial Collagens, Laminin and Fibronectin in vitro

Interaction of human malignant melanoma tumor spheroids with endothelium and reconstituted basement membrane: Modulation by RGDS

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