Changes in the transferrin requirement of cultured chick embryo mesoderm cells during early differentiation

Development ◽  
1986 ◽  
Vol 95 (1) ◽  
pp. 81-93
Author(s):  
E. J. Sanders
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Specific Effect ◽  
Primitive Streak ◽  
Cellular Adhesion ◽  
Type I ◽  
Surface Binding ◽  
Surface Receptors ◽  
Early Differentiation ◽  
Species Specific

Mesodermal tissue from the chick embryo at various stages of early differentiation was cultured in hydrated gels of type I collagen in the presence and absence of transferrin. Primary mesoderm explants from primitive-streak-stage embryos responded to the presence of avian transferrin by significantly improved outgrowth which appeared to be related to the ability of the cells to attach to, and migrate in, the collagen. No evidence was obtained which suggested that this observation was dependent on increased cell proliferation. This outgrowth enhancement was not duplicated by transferrin of human origin. The avian transferrin did not produce this effect on cells cultured on plastic substrata, suggesting that the species-specific effect involves modulation by the extracellular matrix. Mesoderm explants from somite stages of development showed no increase in outgrowth in the presence of either avian or human transferrin as judged by counting the number of outwandering cells. Ultrastructural immunocytochemistry indicated surface binding of transferrin by cells in the gels, and the presence of endogenous transferrin on the surfaces of mesoderm cells in situ and in their extracellular environment. It is suggested that by binding to cell surface receptors, transferrin may be able to influence the strength of cellular adhesion to collagen and hence the capacity for cell locomotion.

scholarly journals Biomimetic Properties of Force-Spun PHBV Membranes Functionalised with Collagen as Substrates for Biomedical Application

Coatings ◽  
2019 ◽  
Vol 9 (6) ◽  
pp. 350 ◽  
Author(s):  
Kegan McColgan-Bannon ◽  
Sarah Upson ◽  
Piergiorgio Gentile ◽  
Muhammad Tausif ◽  
Stephen Russell ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Biomedical Application ◽  
Cell Metabolism ◽  
Polymer Concentration ◽  
Cell Activity ◽  
Cellular Adhesion ◽  
Dermal Fibroblasts ◽  
Type I ◽  
Appreciable Change ◽  
Adhesion Properties

The force-spinning process parameters (i.e., spin speed, spinneret-collector distance, and polymer concentration), optimised and characterised in previous work by this group, allowed the rapid fabrication of large quantities of high surface area poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) polymeric fibre membranes. This paper examined the potential application for force-spun PHBV fibres functionalised with type I collagen for tissue regeneration applications. PHBV fibre scaffolds provide a biologically suitable substrate to guide the regeneration of dermal tissues, however, have poor cellular adhesion properties. The grafting of collagen type-I to PHBV fibres demonstrated improved cell adhesion and growth in Neo-NHDF (neonatal human dermal fibroblasts) fibroblasts. The examination of fibre morphology, thermal properties, collagen content, and degradability was used to contrast the physicochemical properties of the PHBV and PHBV-Collagen fibres. Biodegradation models using phosphate buffered saline determined there was no appreciable change in mass over the course of 6 weeks; a Sirius Red assay was performed on degraded samples, showing no change in the quantity of collagen. Cell metabolism studies showed an increase in cell metabolism on conjugated samples after three and 7 days. In addition, in vitro cytocompatibility studies demonstrated superior cell activity and adhesion on conjugated samples over 7 days.

Measurement of Cellular Adhesion Ratios Under Shear Flow on Various Substrates

2011 ◽  
Author(s):  
Ryo Shirakashi ◽  
Kiyoshi Takano ◽  
Christophe Provin ◽  
Yasuyuki Sakai ◽  
Teruo Fujii
Keyword(s):  
Tissue Culture ◽  
Shear Stress ◽  
Type I Collagen ◽  
Hepatoma Cell ◽  
Glass Plate ◽  
Perfusion Culture ◽  
Cellular Adhesion ◽  
Hepatoma Cell Line ◽  
Type I ◽  
Polystyrene Plate

Perfusion culture is an effective method to enhance the oxygen and nutrient mass transfer for the culture of highly metabolic cells and/or the culture at a high cell density. However, the flow rate of culture medium induces a shear stress that may lead to the death of cells if it is too high. In this study, we measured the cellular adhesion ratio on various materials coated with type-I collagen under Poiseuille flow with flow rates in the range 1–21 mL/min. Hepatoma cell line, HepG2 cells, attached better on a polystyrene plate for tissue culture coated with type-I collagen (with τ0.5, the shear stress required to detach 50% of cells, equal to 42.2 Pa) followed by a collagen coated glass plate (τ0.5 of 40.5Pa), then a polystyrene plate for tissue culture without collagen coating (τ0.5 of 33.8Pa), and finally on a PDMS (τ0.5 of 24.8Pa) plate coated with collagen. The fluorescence staining of the collagen suggests that clumps of cells and collagen were detached from the surface, which implies that the cell-collagen bonds are stronger than collagen-substrate bonds. Accounting these results, it can be concluded that by reinforcing the bonds between collagen and substrate, it might be possible for the cellular monolayer to stay attached on the substrate until τ0.5 reaches ∼40Pa. This conclusion suggests the importance of carefully choosing the cell substrate, which has a strong binding with the coated extracellular matrix, for the cell culture under a high shear stress.

scholarly journals Arg-Gly-Asp-containing peptides expose novel collagen receptors on fibroblasts: implications for wound healing.

1991 ◽  
Vol 2 (12) ◽  
pp. 1035-1044 ◽  
Author(s):  
M V Agrez ◽  
R C Bates ◽  
A W Boyd ◽  
G F Burns
Keyword(s):  
Extracellular Matrix ◽  
Wound Healing ◽  
Type I Collagen ◽  
Regulatory Control ◽  
Type I ◽  
Collagen Gels ◽  
Collagen Matrices ◽  
Surface Receptors ◽  
Rgd Sequence ◽  
Collagen Receptors

Integrins are a family of cell-surface receptors intimately involved in the interactions of cells with their extracellular matrix. These receptors comprise an alpha and beta subunit in noncovalent association and many have been shown to recognize and bind an arginine-glycine-aspartate (RGD) sequence contained within their specific extracellular matrix ligand. Fibroblasts express integrin receptors belonging to two major subfamilies. Some of the members within the subfamily defined by beta 1 (VLA) are receptors for collagen but, perhaps surprisingly, the other major subfamily of integrins on fibroblasts--that defined by the alpha chain of the vitronectin receptor, alpha v--all appear to bind primarily vitronectin and/or fibronectin. In the present study we show that RGD-containing peptides expose cryptic binding sites on the alpha v-associated integrins enabling them to function as collagen receptors. The addition of RGD-containing peptides to fibroblasts cultured on type I collagen induced dramatic cell elongation and, when the cells were contained within collagen matrices, the peptides induced marked contraction of the gels. These processes were inhibited by Fab fragments of a monoclonal antibody against an alpha v integrin. Also, alpha v-associated integrins from cell lysates bound to collagen I affinity columns in the presence, but not in the absence, of RGD-containing peptides. These data suggest a novel regulatory control for integrin function. In addition, because the cryptic collagen receptors were shown to be implicated in the contraction of collagen gels, the generation of such binding forces suggests that this may be the major biological role for these integrins in processes such as wound healing.

scholarly journals Spatiotemporal regulation of PEDF signaling by type I collagen remodeling

2020 ◽  
Vol 117 (21) ◽  
pp. 11450-11458
Author(s):  
Kazuki Kawahara ◽  
Takuya Yoshida ◽  
Takahiro Maruno ◽  
Hiroya Oki ◽  
Tadayasu Ohkubo ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Type I Collagen ◽  
Pigment Epithelium ◽  
Complex Structure ◽  
Binding Activity ◽  
Type I ◽  
Surface Receptors ◽  
Collagen Remodeling ◽  
Collagen Peptide ◽  
Binding Surface

Dynamic remodeling of the extracellular matrix affects many cellular processes, either directly or indirectly, through the regulation of soluble ligands; however, the mechanistic details of this process remain largely unknown. Here we propose that type I collagen remodeling regulates the receptor-binding activity of pigment epithelium-derived factor (PEDF), a widely expressed secreted glycoprotein that has multiple important biological functions in tissue and organ homeostasis. We determined the crystal structure of PEDF in complex with a disulfide cross-linked heterotrimeric collagen peptide, in which the α(I) chain segments—each containing the respective PEDF-binding region (residues 930 to 938)—are assembled with an α2α1α1 staggered configuration. The complex structure revealed that PEDF specifically interacts with a unique amphiphilic sequence, KGHRGFSGL, of the type I collagen α1 chain, with its proposed receptor-binding sites buried extensively. Molecular docking demonstrated that the PEDF-binding surface of type I collagen contains the cross-link–susceptible Lys930 residue of the α1 chain and provides a good foothold for stable docking with the α1(I) N-telopeptide of an adjacent triple helix in the fibril. Therefore, the binding surface is completely inaccessible if intermolecular crosslinking between two crosslink-susceptible lysyl residues, Lys9 in the N-telopeptide and Lys930, is present. These structural analyses demonstrate that PEDF molecules, once sequestered around newly synthesized pericellular collagen fibrils, are gradually liberated as collagen crosslinking increases, making them accessible for interaction with their target cell surface receptors in a spatiotemporally regulated manner.

scholarly journals Effect of L-azetidine-2-carboxylic acid on glycosylations of collagen in chick-embryo tendon cells

1976 ◽  
Vol 160 (3) ◽  
pp. 639-645 ◽  
Author(s):  
A Oikarinen ◽  
H Anttinen ◽  
K I Kivirikko
Keyword(s):  
Carboxylic Acid ◽  
Chick Embryo ◽  
Type I Collagen ◽  
Type I ◽  
Pulse Label ◽  
Chase Period ◽  
Helix Formation ◽  
Collagen Secretion ◽  
Tendon Cells ◽  
Triple Helix Formation

The glycosylations of hydroxylysine during collagen biosynthesis in isolated chick-embryo tendon cells were studied by using pulse-chase labelling experiments with [14C]-lysine. The hydroxylation of lysine and the glycosylations of hydroxylysine continued after a 5 min pulse label for up to about 10 min during the chase period. These data differ from those obtained previously in isolated chick-embryo cartilage cells, in which, after a similar 5 min pulse label, these reactions continued during the chase period for up to about 20 min. The collagen synthesized by the isolated chick-embryo tendon cells differed markedly from the type I collagen of adult tissues in its degree of hydroxylation of lysine residues and glycosylations of hydroxylysine residues. When the isolated tendon cells were incubated in the presence of L-azetidine-2-carboxylic acid, the degree of glycosylations of hydroxylysine during the first 10 min of the chase period was identical with that in cells incubated without thcarboxylic acid for at least 60 min, whereas no additional glycosylations took place in the control cells after the 10 min time-point. As a consequence, the collagen synthesized in the presence of this compound contained more carbohydrate than did the collagen synthesized by the control cells. Additional experiments indicated that azetidine-2-carboxylic acid did not increase the collagen glycosyltransferase activities in the tendon cells or the rate of glycosylation reactions when added directly to the enzyme incubation mixture. Control experiments with colchicine indicated that the delay in the rate of collagen secretion, which was observed in the presence of azetidine-2-carboxylic acid, did not in itself affect the degree of glycosylations of collagen. The results thus suggest that the increased glycosylations were due to inhibition of the collagen triple-helix formation, which is known to occur in the presence of azetidine-2-carboxylic acid.

scholarly journals Binding of soluble type I collagen to fibroblasts: specificities for native collagen types, triple helical structure, telopeptides, propeptides, and cyanogen bromide-derived peptides.

1982 ◽  
Vol 95 (3) ◽  
pp. 752-756 ◽  
Author(s):  
B D Goldberg ◽  
R E Burgeson
Keyword(s):  
Cyanogen Bromide ◽  
Type I Collagen ◽  
Helical Structure ◽  
Type I ◽  
Collagen Types ◽  
Surface Receptors ◽  
Type I Procollagen ◽  
Competitive Inhibitors ◽  
A Minor

Unlabeled collagenous proteins were quantified as inhibitors of binding of native, soluble, radioiodinated type I collagen to the fibroblast surface. Collagen types IV, V a minor cartilage isotype (1 alpha 2 alpha 3 alpha), and the collagenlike tail of acetylcholinesterase did not inhibit binding. Collagen types II and III behaved as competitive inhibitors of type I binding. Denaturation of native collagenous molecules exposed cryptic inhibitory determinants in the separated constituent alpha chains. Inhibition of binding by unlabeled type I collagen was not changed by enzymatic removal of the telopeptides. Inhibitory determinants were detected in cyanogen bromide-derived peptides from various regions of helical alpha 1 (I) and alpha 1(III) chains. The aminoterminal propeptide of chick pro alpha 1(I) was inhibitory for binding, whereas the carboxyterminal three-chain propeptide fragment of human type I procollagen was not. The data are discussed in terms of the proposal that binding to surface receptors initiates the assembly of periodic collagen fibrils in vivo.

scholarly journals Catalytic properties of lysyl hydroxylase from cells synthesizing genetically different collagen types

1982 ◽  
Vol 201 (1) ◽  
pp. 215-219 ◽  
Author(s):  
U Puistola
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Collagen Types ◽  
Enzyme Preparations ◽  
Lysyl Hydroxylase ◽  
Type Iv ◽  
Sarcoma Cells

Crude preparations of lysyl hydroxylase were extracted from chick-embryo tendons synthesizing exclusively type I collagen, chick-embryo sterna synthesizing exclusively type II collagen and HT-1080 sarcoma cells synthesizing exclusively type IV collagen. No differences were found in the Km values for Fe2+, 2-oxoglutarate and ascorbate between these three enzymes preparations. Similarly no differences were found in the Km values for type I and type II protocollagens and the rate at which type IV protocollagen is hydroxylated between these enzyme preparations. The extent to which type I protocollagen could be hydroxylated by the three enzymes was likewise identical. These data strongly argue against the existence of collagen-type-specific lysyl hydroxylase isoenzymes.

scholarly journals Hyaluronan Cell Surface Binding Is Induced by Type I Collagen and Regulated by Caveolae in Glioma Cells

2004 ◽  
Vol 279 (21) ◽  
pp. 21888-21896 ◽  
Author(s):  
Borhane Annabi ◽  
Sébastien Thibeault ◽  
Robert Moumdjian ◽  
Richard Béliveau
Keyword(s):  
Cell Surface ◽  
Type I Collagen ◽  
Glioma Cells ◽  
Type I ◽  
Surface Binding ◽  
Cell Surface Binding

scholarly journals Inhibition of prolyl hydroxylation and procollagen processing in chick-embryo calvaria by a derivative of pyridine-2,4-dicarboxylate. Characterization of the diethyl ester as a proinhibitor

1991 ◽  
Vol 275 (2) ◽  
pp. 469-476 ◽  
Author(s):  
G Tschank ◽  
D G Brocks ◽  
K Engelbart ◽  
J Mohr ◽  
E Baader ◽  
...  
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Helical Structure ◽  
Diethyl Ester ◽  
Type I ◽  
Dependent Manner ◽  
Maximal Inhibition ◽  
Collagen Formation ◽  
Collagenous Material

The biochemical and morphological consequences of procollagen prolyl 4-hydroxylase inhibition by pyridine-2,4-dicarboxylic acid (2,4-PDCA) and its diethyl ester (diethyl-2,4-PDC) were studied in chick-embryo calvaria, which predominantly synthesize type I collagen. Half-maximal inhibition of tissue hydroxyproline formation required 650 microM-2,4-PDCA, whereas the Ki with respect to chicken prolyl 4-hydroxylase in vitro was 2 microM. In contrast, half-maximal inhibition was caused by 10 microM-diethyl-2,4-PDC in the intact calvaria, although chicken prolyl 4-hydroxylase in vitro was not inhibited even at 1 mM. The collagenous material produced in the presence of diethyl-2,4-PDC showed an altered ‘melting’ profile and a lowering of the transition temperature by 10 degrees C, indicating misalignment and thermal instability of its triple-helical structure. Amount and electrophoretic mobility of procollagen type I chains were increased in a dose-dependent manner. The amounts of partially processed species and alpha-chains were decreased, without change in mobility. This marked effect on procollagen-collagen conversion in the intact calvaria suggests that the underhydroxylated collagenous material generated in the presence of diethyl-2,4-PDC is resistant to or acts as endogenous secondary inhibitor of type I procollagen N-proteinase. Electron microscopy of treated calvaria cells showed dilated rough endoplasmic reticulum and numerous phagolysosomes, indicating intracellular retention and lysosomal degradation of the newly synthesized underhydroxylated collagenous material. In summary, these results identify 2,4-PDCA and diethyl-2,4-PDC as the first prolyl 4-hydroxylase-directed inhibitor/proinhibitor pair that affects intra- and extra-cellular events during collagen formation.

scholarly journals Inhibition of fibronectin binding and fibronectin-mediated cell adhesion to collagen by a peptide from the second type I repeat of thrombospondin.

1993 ◽  
Vol 121 (2) ◽  
pp. 469-477 ◽  
Author(s):  
J M Sipes ◽  
N Guo ◽  
E Nègre ◽  
T Vogel ◽  
H C Krutzsch ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Adhesion ◽  
Type I Collagen ◽  
Selective Inhibitor ◽  
Type I ◽  
Surface Receptors ◽  
Extracellular Matrix Components ◽  
Flanking Sequences ◽  
Fibronectin Binding

The platelet and extracellular matrix glycoprotein thrombospondin interacts with various types of cells as both a positive and negative modulator of cell adhesion, motility, and proliferation. These effects may be mediated by binding of thrombospondin to cell surface receptors or indirectly by binding to other extracellular matrix components. The role of peptide sequences from the type I repeats of thrombospondin in its interaction with fibronectin were investigated. Fibronectin bound specifically to the peptide Gly-Gly-Trp-Ser-His-Trp from the second type I repeat of thrombospondin but not to the corresponding peptides from the first or third repeats or flanking sequences from the second repeat. The two Trp residues and the His residue were essential for binding, and the two Gly residues enhanced the affinity of binding. Binding of the peptide and intact thrombospondin to fibronectin were inhibited by the gelatin-binding domain of fibronectin. The peptide specifically inhibited binding of fibronectin to gelatin or type I collagen and inhibited fibronectin-mediated adhesion of breast carcinoma and melanoma cells to gelatin or type I collagen substrates but not direct adhesion of the cells to fibronectin, which was inhibited by the peptide Gly-Arg-Gly-Asp-Ser. Thus, the fibronectin-binding thrombospondin peptide Gly-Gly-Trp-Ser-His-Trp is a selective inhibitor of fibronectin-mediated interactions of cells with collagen in the extracellular matrix.

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