An endothelial cell growth factor derived from human lung carcinoma cells grown in serum-free medium

Walker, Carol; Mates, G.; Pumford, D.; Daniel, M.R.

doi:10.1242/jcs.87.5.739

Cited by 4 publications

(2 citation statements)

References 36 publications

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“…The observed biological activity of VEGF released from VBP and VBP WT microspheres agrees with previous observations that released VEGF acts as an endothelial cell mitogen . Importantly, serum concentration alone did not influence HUVEC proliferation (Figure ) in agreement with previous literature, , which enabled us to specifically study the impact of serum concentration on VEGF activity in vitro . The biological activity of released VEGF in serum-containing medium (Figure ) is consistent with our observation that VEGF release is not influenced strongly by protease activity in a high concentration of serum (Figure ).…”

Section: Discussionsupporting

confidence: 91%

Serum-Dependence of Affinity-Mediated VEGF Release from Biomimetic Microspheres

et al. 2014

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Vascular endothelial growth factor (VEGF) activity is highly regulated via sequestering within the ECM and cell-demanded proteolysis to release the sequestered VEGF. Numerous studies have demonstrated that VEGF activity mediates cellular events leading to angiogenesis and capillary formation in vivo. This has motivated the study of biomaterials to sustain VEGF release, and in many cases, the materials are inspired by the structure and function of the native ECM. However, there remains a need for materials that can bind to VEGF with high specificity, as the in vivo environment is rich in a variety of growth factors (GFs) and GF-binding moieties. Here we describe a strategy to control VEGF release using hydrogel microspheres with tethered peptides derived from VEGF receptor 2 (VEGFR2). Using biomaterials covalently modified with varying concentrations of two distinct VEGFR2-derived peptides with varying serum stability, we analyzed both biomaterial and environmental variables that influence VEGF release and activity. The presence of tethered VEGF-binding peptides (VBPs) resulted in significantly extended VEGF release relative to control conditions, and the resulting released VEGF significantly increased the expansion of human umbilical vein endothelial cells in culture. VEGF release rates were also strongly influenced by the concentration of serum. The presence of Feline McDonough Sarcoma-like tyrosine kinase 1 (sFlt-1), a serum-borne receptor fragment derived from VEGF receptor 1, increased VEGF release rates, although sFlt-1 was not sufficient to recapitulate the release profile of VEGF in serum. Further, the influence of serum on VEGF release was not due to protease activity or nonspecific VEGF interactions in the presence of serum-borne heparin. VEGF release kinetics correlated well with a generalizable mathematical model describing affinity-mediated release of VEGF from hydrogel microspheres in defined conditions. Modeling results suggest a potential mechanism whereby competition between VEGF and multiple VEGF-binding serum proteins including sFlt-1, soluble kinase insert domain receptor (sKDR), and α2-macroglobulin (α2-M) likely influenced VEGF release from microspheres. The materials and mathematical model described in this approach may be useful in a range of applications in which sustained, biologically active GF release of a specific GF is desirable.

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Section: Discussionsupporting

confidence: 91%

Serum-Dependence of Affinity-Mediated VEGF Release from Biomimetic Microspheres

et al. 2014

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“…Our results indicate that platelets contain a potent endothelial cell mitogen that is distinct from FGFs. Recently, an endothelial cell mitogen which, like PD-ECGF, does not have affinity for heparin was found to be produced by a poorly differentiated bronchial carcioma cell line, 1PT (Walker et al, 1987). The 1PT cell-derived growth factor has similar biological and biochemical properties as PD-ECGF; whether it is identical with PD-ECGF remains to be elucidated.…”

Section: Discussionmentioning

confidence: 99%

High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum

Miyazono¹,

Heldin²

1989

Biochemistry

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Platelet-derived endothelial cell growth factor (PD-ECGF) is a 45-kDa protein that stimulates the growth of endothelial cells [Miyazono, K., et al. (1987) J. Biol. Chem. 262, 4098-4103]. Here, we describe a method to purify large quantities of PD-ECGF from human platelet lysate at a high yield (14% overall recovery). The purification method involves five steps, using high-performance liquid chromatography grade hydroxylapatite and hydrophobic chromatographies as the two final steps. The purified material contained two major components of apparent molecular weight values of 46,000 and 44,000. These components coeluted in a high-resolving reversed-phase chromatography and were found to give similar peptide maps after treatments with staphylococcal V8 protease, suggesting that the 44-kDa form is related to the 46-kDa molecule. Partial tryptic digestion of native PD-ECGF revealed that the molecule contains a trypsin-resistant domain of 37-39 kDa. A rabbit antiserum was produced against the purified material and was found to specifically recognize PD-ECGF in immunoblotting. When added to the cell culture medium, an immunoglobulin fraction of the antiserum neutralized the activity of purified PD-ECGF. Furthermore, it completely neutralized the endothelial cell mitogenic activity of platelet lysate, indicating that PD-ECGF is the only mitogen in platelet lysate for this cell type.

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Characteristics of two cell lines derived from a histologically undifferentiated bronchial carcinoma expressing neuroendocrine markers

Wright-Perkins

Daniel

Walker

1994

In Vitro Cell Dev Biol - Animal

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This study investigates the characteristics of two human cell lines--1PT and 1PT VARIANT A--both derived from the same histologically undifferentiated, neuroendocrine positive, non-small cell lung carcinoma (NSCLC) and capable of growth in unsupplemented serum-free minimum essential medium. In stationary culture, the cells of both lines grew both attached to a plastic substratum and in suspension; the 1PT VARIANT A line formed three-dimensional clusters of loosely adherent cells. The cell lines differed in their DNA content, the 1PT having 1.44 times and the 1PT VARIANT A having 2.39 times the normal human diploid DNA content. Chromosome counts supported this observation, the ploidy of the 1PT and VARIANT A lines being 1.11 and 1.64, respectively. On transmission electron microscopy the cells of both lines had dense core granules and immature desmosomes, whereas only the 1PT VARIANT A line had mucin granules. Both lines formed, in nude mice, tumors that, like the original tumor from which they were derived, were histologically undifferentiated and showed local invasion. The original tumor and both lines had demonstrable neuroendocrine markers. Cytokeratins were apparent in the tumor but not the cell lines, and neurofilaments were present in the cell lines only. Staining for epithelial membrane antigen, neural cell adhesion molecule, and desmoplakin differentiated between the two lines. These lines provide a useful model for the investigation of the biology of the neuroendocrine positive subgroup of NSCLC, which is clinically important because of the possible responsiveness of these tumors to chemotherapy.

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An endothelial cell growth factor derived from human lung carcinoma cells grown in serum-free medium

Cited by 4 publications

References 36 publications

Serum-Dependence of Affinity-Mediated VEGF Release from Biomimetic Microspheres

Serum-Dependence of Affinity-Mediated VEGF Release from Biomimetic Microspheres

High-yield purification of platelet-derived endothelial cell growth factor: structural characterization and establishment of a specific antiserum

Characteristics of two cell lines derived from a histologically undifferentiated bronchial carcinoma expressing neuroendocrine markers

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