Plasma Fibronectin: Three Steps to Purification and Stability

Poulouin, Laurent; Gallet, Olivier; Rouahi, Myriam; Imhoff, Jean-Marie

doi:10.1006/prep.1999.1103

Cited by 88 publications

(48 citation statements)

References 26 publications

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“…Blocking anti-Vn (MAb1945) was purchased from Chemicon (Hampshire, United Kingdom). Vn and fibronectin (Fn) were purified from cryoprecipitated human plasma as previously described [24,25].…”

Section: Methodsmentioning

confidence: 99%

Initial formation of IGROV1 ovarian cancer multicellular aggregates involves vitronectin

et al. 2010

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Ovarian cancer progression is frequently associated with the development of malignant ascites. Multicellular aggregates of carcinoma cells (spheroids) found within ascites are thought to be able to promote peritoneal carcinomatosis. We have previously demonstrated the involvement of the vitronectin/alphav integrin adhesive system in the dissemination of ovarian cancer cells and continue to investigate the influence of these molecules by studying their role(s) in spheroid behavior. The aim of this study was to generate ovarian cancer multicellular aggregates and to focus on the role of vitronectin and alphav integrins in their initiation. IGROV1 cancer cells cultured in the absence of adhesive substratum formed multicellular aggregates comparable to spheroids. After 21 days, a fraction of the cells within clusters remained viable and proliferated recurrently. Within the multicellular aggregates, vitronectin and alphav integrins were co-localized at intercellular sites, suggesting their involvement in cell-cell interactions. Initial formation of IGROV1 aggregates was inhibited using anti-vitronectin and anti-alphav integrin blocking antibodies or the cyclic peptide cRGDfV. Vitronectin expression persisted during cluster disaggregation on fibronectin. These results demonstrate the ability of IGROV1 cells to generate multicellular aggregates and point to a contributory role for the vitronectin/alphav integrin system in the initial step of this process. These events could represent a prerequisite for further dissemination.

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Section: Methodsmentioning

confidence: 99%

Initial formation of IGROV1 ovarian cancer multicellular aggregates involves vitronectin

et al. 2010

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“…[25] FN purified from adult human plasma by a protocol adapted from Poulouin et al was diluted to a final concentration of 20 mg Á mL À1 in phosphate buffered saline (PBS) (Dulbecco's PBS, Fisher Scientific, Schwerte, Germany). [26] The PEMA-coated microstructures were then incubated in FN solution for 1 h and subsequently rinsed several times with PBS. COLI (PureCol, Nutacon BV, Leimuiden, Netherlands) was immobilized at a final concentration of 0.5 mg Á mL À1 (on ice, 8 parts PureCol, 1 part 10 Â NaOH, 1 part 10 Â PBS and diluted to 1 mg Á mL À1 with the respective amount of PBS at pH 7.4) for 3 h and rinsed several times in MilliQ water.…”

Section: Surface Functionalizationmentioning

confidence: 99%

Extracellular Matrix Functionalized Microcavities to Control Hematopoietic Stem and Progenitor Cell Fate

Kurth

Franke

Pompe

et al. 2011

Macromolecular Bioscience

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Polymeric microcavities functionalized with extracellular matrix components were used as an experimental in vitro model to investigate principles of hematopoietic stem and progenitor cell (HSPC) fate control. Using human CD133+ HSPC we could demonstrate distinct differences in HSPC cycling and differentiation dependence on the adhesion ligand specificity (i.e., heparin, collagen I) and cytokine levels. The presented microcavity platform provides a powerful in vitro approach to explore the role of exogenous cues in HSPC fate decisions and can therefore be instrumental to progress in stem cell biology and translational research toward new therapies.

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“…Plates coated with Basal-ECM, RPMI-ECM or CmOV1-ECM were prepared as described above. Plates were also coated overnight at 4°C with either cellular Fn (cFn, 10 g/mL), laboratory purified Fn (pFn, 1 g/mL), 19 vitronectin (Vn, 10 g/mL), collagen type I (Coll, 125 g/mL), laminin (LM, 20 g/mL), ECM gel (3-24 g/mL), poly-lysine (PL). The concentrations of ECM proteins chosen were optimal.…”

Section: Cell Adhesion Assaymentioning

confidence: 99%

Involvement of αvβ 3 integrin and disruption of endothelial fibronectin network during the adhesion of the human ovarian adenocarcinoma cell line IGROV1 on the human umbilical vein cell extracellular matrix

Carreiras

Thiébot

Leroy-Dudal³

et al. 2002

Intl Journal of Cancer

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Key words: extracellular matrix; integrin; ovarian carcinoma; HUVECsNeoangiogenesis is the complex process by which new blood vessels are generated from preexisting vessels. The growth of new blood vessels plays a critical role in a variety of normal physiological events but also in pathological processes. 1 In the case of tumoral angiogenesis, the process is under cell tumor influence and conversely the expansion of solid tumors is critically dependent on vascular networks that provide nutrients for growing tumors and allow metastasis in distant organs. 2 The interaction of tumors with the vasculature is an important step in the metastatic cascade. The identification of molecules that play a functional role in tumor neovascularization, especially in the interactions of tumor cells with endothelial extracellular matrix underlining the vasculature, has been investigated for several years and the potential role of cell adhesion molecules and their ligands are of interest. 2 The integrins represent the main extracellular matrix (ECM) receptors that are composed of ␣ and ␤ chains that combine to give 23 heterodimers with distinct cellular and adhesive specificities. 3,4 These transmembrane glycoproteins are expressed by endothelial cells as well as by a variety of tumor cells. It was shown that integrin ␣v␤3 can provide specific cell survival signals that facilitate vascular cell proliferation during angiogenesis. 5 This integrin appeared also to be strongly involved in several tumor cell growth such as melanoma and breast cancer. 6,7 ␣v␤3 is a receptor for a wide variety of ECM proteins; among them, fibronectin (Fn) has been involved in tumor development and angiogenesis. 8 Indeed, endothelial cell motility is promoted by binding ECM proteins such as Fn to integrins and interfering with this step may inhibit the process of angiogenesis. 9 Ovarian carcinoma is one of the most common fatal gynecological malignancies and for the majority of patients, these tumors are diagnosed at advanced stages. 10 The ovarian tumors arise mainly from the malignant transformation of ovarian surface epithelium (OSE) cells. These cells spread out and disseminate into the peritoneal cavity to give intra-abdominal grafts whereas metastases in distant organs occurred late. There are only few reports regarding angiogenesis in patients with advanced ovarian carcinoma. Quantification and evaluation of vascular density in ovarian cancer revealed that tumor angiogenesis is a prognostic factor. 11,12 Little is known, however, about the cellular processes that lead to ovarian carcinoma neovascularization. Because several studies demonstrated the primordial role of integrins in ovarian cancer behavior, in particular of ␣v␤3 integrin and their ligands, 13-15 we focused our interest on the role of this integrin in the interactions between ovarian tumor and its vasculature.In our study, we therefore used in vitro models, IGROV1 human ovarian adenocarcinoma cell line and human umbilical vein endothelial cells (HUVECs) to analyze the adhesion properti...

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Plasma Fibronectin: Three Steps to Purification and Stability

Cited by 88 publications

References 26 publications

Initial formation of IGROV1 ovarian cancer multicellular aggregates involves vitronectin

Initial formation of IGROV1 ovarian cancer multicellular aggregates involves vitronectin

Extracellular Matrix Functionalized Microcavities to Control Hematopoietic Stem and Progenitor Cell Fate

Involvement of αvβ 3 integrin and disruption of endothelial fibronectin network during the adhesion of the human ovarian adenocarcinoma cell line IGROV1 on the human umbilical vein cell extracellular matrix

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