scholarly journals Basic fibroblast growth factor promotes adhesive interactions of neuroepithelial cells from chick neural tube with extracellular matrix proteins in culture

Development ◽  
1993 ◽  
Vol 119 (3) ◽  
pp. 943-956 ◽  
Author(s):  
Y. Kinoshita ◽  
C. Kinoshita ◽  
J.G. Heuer ◽  
M. Bothwell
Keyword(s):  
Extracellular Matrix ◽  
Neural Tube ◽  
Cell Spreading ◽  
Matrix Proteins ◽  
Fibroblast Growth ◽  
Spreading Effect ◽  
Neuroepithelial Cells ◽  
Beta 1 Integrin ◽  
Adhesive Interactions ◽  
Cell Plating

Fibroblast growth factors have been increasingly assigned mitogenic and trophic roles in embryonic and postnatal development of the nervous system. Little is known, however, of their functional roles in early embryonic neural development at the neural tube stage. We have examined the effect of basic fibroblast growth factor (bFGF) on the adhesive behavior in culture of dissociated brachio-thoracic neural tube cells from 26- to 30-somite stage chick embryos. Cells plated on collagen-coated substratum at a low density attach to the substratum but show poor cell spreading. Addition of bFGF markedly promotes cell spreading, yielding an epithelial morphology. This effect becomes discernible 6–8 hours after cell plating with bFGF and is completed by 24 hours, with half-maximal and maximal effects attained at around 0.4 and 10 ng/ml, respectively. The number of cells remain largely constant up to 24 hours, and then cell survival and/or mitogenic effects of bFGF become apparent. The cell spreading effect is abolished by cycloheximide treatment, inhibited by the anti-beta 1-integrin antibody CSAT, and accompanied by about twofold increases in the expression of beta 1-integrin and vinculin, components of focal adhesion complexes. Cells cultured with bFGF for 24 hours exhibit enhanced cell attachment and cell spreading with little time lag following cell plating. In earlier embryonic stages, developmentally less mature cells depend much more on bFGF for their cell spreading and survival, while in later stages the cell spreading response to bFGF becomes undetectable as neural tube develops to spinal cord. The cell spreading effect of bFGF is realized on specific extracellular matrix proteins including laminin, fibronectin and collagen, but not on vitronectin, arg-gly-asp peptide (PepTite-2000), poly-L-ornithine or others. These results suggest that, in an early stage of neural tube development, bFGF is involved in the developmental regulation of adhesive interactions between neuroepithelial cells and the extracellular matrix, thereby controlling their proliferation, migration and differentiation.

Induction of Hepatocyte Differentiation by the Extracellular Matrix and an RGD-Containing Synthetic Peptide

1991 ◽  
Vol 252 ◽  
Author(s):  
David J. Mooney ◽  
Robert Langer ◽  
Linda K. Hansen ◽  
Joseph P. Vacanti ◽  
Donald E. Ingber
Keyword(s):  
Extracellular Matrix ◽  
Protein Secretion ◽  
Synthetic Peptide ◽  
Cell Spreading ◽  
Specific Protein ◽  
Hepatocyte Differentiation ◽  
Liver Specific Protein ◽  
Extensive Cell ◽  
Adhesive Interactions

ABSTRACTTo design novel biomaterials for hepatocyte transplantation it will be necessary to determine whether specific extracellular matrix (ECM) molecule(s) or the adhesive interactions between the surface and hepatocytes are responsible for regulation of hepatocyte function. Purified ECM molecules (laminin, fibronectin, types I and IV collagen) and a synthetic peptide containing the arginine-glycine-aspartate (RGD) cell-binding sequence were precoated at defined densities to non-adhesive polystyrene dishes. Hepatocytes cultured on dishes coated with a low density of ECM molecules (1 ng/cm2) maintained a round morphology, and high liver-specific protein secretion rates. In contrast, culturing hepatocytes on increasing ECM densities (50–1000 ng/cm2) resulted in extensive cell spreading, a loss of liver-specific protein secretion, and cell growth. Hepatocytes cultured on dishes coated with the RGD-containing peptide did not spread even on a high density of the peptide (10,000 ng/cm2), and albumin secretion remained high for hepatocytes cultured on all peptide densities (1–10,000 ng/cm2). These results suggest that a variety of ECM molecules and synthetic peptides are capable of inducing hepatocyte differentiation in vitro, and these effects depend on their ability to promote cell spreading.

Receptor-mediated adhesive and anti-adhesive functions of chondroitin sulfate proteoglycan preparations from embryonic chicken brain

1995 ◽  
Vol 108 (12) ◽  
pp. 3807-3816 ◽  
Author(s):  
H. Ernst ◽  
M.K. Zanin ◽  
D. Everman ◽  
S. Hoffman
Keyword(s):  
Extracellular Matrix ◽  
Chondroitin Sulfate ◽  
Cell Attachment ◽  
Extracellular Matrix Proteins ◽  
Cell Spreading ◽  
The Other ◽  
Matrix Proteins ◽  
Surface Receptors ◽  
Core Proteins ◽  
Proteoglycan Aggregates

Chondroitin sulfate proteoglycans inhibit the adhesion of cells to extracellular matrix proteins that otherwise permit adhesion. Although proteoglycans are widely assumed to act by masking the other protein in a mixed substrate, recent studies suggest that proteoglycans inhibit adhesion through mechanisms initiated by their binding to specific cell surface receptors. To explore this issue, we developed a purification scheme to isolate proteoglycan aggregates, monomers, and core proteins. Two distinct adhesion assays were used to study the interaction of these proteoglycan preparations with human foreskin fibroblasts: the gravity assay in which cell attachment is stabilized by cell spreading, and the centrifugation assay in which spreading does not play a role. All proteoglycan preparations mediate adhesion in the centrifugation assay but not in the gravity assay. In the centrifugation assay, proteoglycan aggregates and monomers are considerably more active than other extracellular matrix proteins while proteoglycan core proteins are at least as active as other extracellular matrix proteins. Proteoglycan core proteins bind to cell-associated hyaluronic acid, but not to integrins. Using mixed substrates in the gravity assay, all proteoglycan preparations inhibited cell attachment to fibronectin and vitronectin but not to collagen I and laminin. Although proteoglycan aggregates and monomers are more active than core proteins in inhibiting adhesion in the gravity assay, core proteins are still clearly active. A variety of control experiments suggest that the inhibition of cell attachment by proteoglycans is mediated through the specific interactions of proteoglycans with cell surface receptors, resulting in the inhibition of cell spreading. These results suggest at least two molecular mechanisms for proteoglycan-fibroblast interactions, one involving the chondroitin sulfate on the proteoglycan and an as yet unidentified receptor, the other involving the proteoglycan core protein and cell-associated hyaluronic acid.

Developmentally regulated expression of alpha 6 integrin in avian embryos

Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 197-211 ◽  
Author(s):  
M. Bronner-Fraser ◽  
M. Artinger ◽  
J. Muschler ◽  
A.F. Horwitz
Keyword(s):  
Nervous System ◽  
Neural Tube ◽  
Integrin Subunit ◽  
Developmentally Regulated ◽  
Commissural Neurons ◽  
Regulated Expression ◽  
Neuroepithelial Cells ◽  
Beta 1 Integrin ◽  
Intense Immunoreactivity ◽  
Developing Nervous System

The distribution pattern of the avian alpha 6 integrin subunit was examined during early stages of development. The results show that this subunit is prevalent in cells of the developing nervous system and muscle. alpha 6 is first observed on neuroepithelial cells of the cranial neural plate and trunk neural tube. With time, immunoreactivity becomes prominent near the lumen and ventrolateral portions of the neural tube, co-distributing with neurons and axons, particularly notable on commissural neurons. The alpha 6 expression pattern is dynamic in the neural tube, with immunoreactivity peaking by embryonic day 6 (stage 30) and decreasing thereafter. The ventral roots and retina exhibit high levels of immunoreactivity throughout development. In the peripheral nervous system, alpha 6 immunoreactivity first appears on a subpopulation of sympathoadrenal cells around the dorsal aorta and later in the dorsal root ganglia shortly after gangliogenesis. Immunoreactivity appears on prospective myotomal cells as the somites delaminate into the dermomyotome and sclerotome, remaining prominent on myoblasts and differentiated muscle at all stages. The mesonephros also has intense immunoreactivity. In the periphery, alpha 6 immunoreactive regions often in proximity to laminin, which is thought to be the ligand of alpha 6 beta 1 integrin.

scholarly journals Adhesion of a chicken myeloblast cell line to fibrinogen and vitronectin through a beta 1-class integrin.

1992 ◽  
Vol 116 (3) ◽  
pp. 809-815 ◽  
Author(s):  
K M Neugebauer ◽  
K A Venstrom ◽  
L F Reichardt
Keyword(s):  
Extracellular Matrix ◽  
Cell Adhesion ◽  
Cell Line ◽  
Cell Attachment ◽  
Matrix Proteins ◽  
Adhesive Properties ◽  
Hd11 Cell ◽  
Adhesive Interactions ◽  
Alpha V Beta 3 ◽  
Blocking Antibodies

The adhesive interactions of circulating blood cells are tightly regulated, receptor-mediated events. To establish a model for studies on regulation of cell adhesion, we have examined the adhesive properties of the HD11 chick myeloblast cell line. Function-perturbing antibodies were used to show that integrins containing the beta 1 subunit mediate HD11 cell attachment to several distinct extracellular matrix proteins, specifically fibronectin, collagen, vitronectin, and fibrinogen. This is the first evidence that an integrin heterodimer in the beta 1 family functions as a receptor for fibrinogen. While the alpha v beta 1 heterodimer has been shown to function as a vitronectin receptor on some cells, this heterodimer could not be detected on HD11 cells. Instead, results suggest that the beta 1 subunit associates with different, unidentified alpha subunit(s) to form receptors for vitronectin and fibrinogen. Results using function-blocking antibodies also demonstrate that on these cells, additional receptors for vitronectin are formed by alpha v beta 3 and alpha v associated with an unidentified 100-kD beta subunit. The adhesive interactions of HD11 cells with these extracellular matrix ligands were shown to be regulated by lipopolysaccharide treatment, making the HD11 cell line attractive for studies of mechanisms regulating cell adhesion. In contrast to primary macrophage which rapidly exhibit enhanced adhesion to laminin and collagen upon activation, activated HD11 cells exhibited reduced adhesion to most extracellular matrix constituents.

Cardiac myocytes cultured on a basement membrane extract of the ehs tumor

1986 ◽  
Vol 44 ◽  
pp. 270-271
Author(s):  
L. Terracio ◽  
A. Dewey ◽  
K. Rubin ◽  
T.K. Borg
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Cardiac Myocytes ◽  
Normal Tissue ◽  
Cell Spreading ◽  
Collagen Iv ◽  
Basement Membrane Extract ◽  
Membrane Extract ◽  
Growth Development ◽  
Multicellular Organisms

The recognition and interaction of cells with the extracellular matrix (ECM) effects the normal physiology as well as the pathology of all multicellular organisms. These interactions have been shown to influence the growth, development, and maintenance of normal tissue function. In previous studies, we have shown that neonatal cardiac myocytes specifically interacts with a variety of ECM components including fibronectin, laminin, and collagens I, III and IV. Culturing neonatal myocytes on laminin and collagen IV induces an increased rate of both cell spreading and sarcomerogenesis.

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

1997 ◽  
Vol 10 (01) ◽  
pp. 6-11 ◽  
Author(s):  
R. F. Rosenbusch ◽  
L. C. Booth ◽  
L. A. Dahlgren
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Tendon Healing ◽  
Matrix Proteins ◽  
Isolated Cells ◽  
Superficial Digital Flexor Tendon ◽  
Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

Extracellular matrix proteins control the growth of cerebellar medulloblastoma cells

2004 ◽  
Vol 216 (03) ◽  
Author(s):  
U Schüller ◽  
W Hartmann ◽  
A Koch ◽  
K Schilling ◽  
OD Wiestler ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Extracellular Matrix Proteins ◽  
Matrix Proteins ◽  
Cerebellar Medulloblastoma
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