Substrate adhesion of rat hepatocytes: on the mechanism of attachment to fibronectin.
Staffan Johansson,Magnus Höök +1 more
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Data is presented supporting a mechanism of cell attachment to fibronectin involving multiple weak interactions between cellular receptors and substrate molecules, although some steps in the attachment process appear to disobey the requirements for such a mechanism.Abstract:
We examined the mechanisms of cell attachment to fibronectin-coated substrates. Inhibition of cell attachment was obtained by species-specific antifibronectin antibodies, which presumably recognize a distinct antigenic structure in the protein located at, or in the immediate vicinity of, the cell-binding site. The inhibiting antibodies could be adsorbed on a column of Sepharose substituted with plasma fibronectin. The initial phase of cell attachment was also inhibited by addition of soluble fibronectin to the incubation medium in a reaction that exhibited specificity and concentration dependence. These data suggest that cell-binding sites are available in an active form on the surface of soluble fibronectin. However, the inhibitory effect of fibronectin was greatly enhanced by adding the protein together with heparin, heparan sulfate, collagen, or a fibronectin-binding collagen peptide (CB-7), which is consistent with an "activation" of fibronectin on binding to these matrix components. A similar activation of fibronectin was obtained by cleaving the protein with trypsin. We discuss these findings in relation to conformational rearrangements in the fibronectin molecule. Data is presented supporting a mechanism of cell attachment to fibronectin involving multiple weak interactions between cellular receptors and substrate molecules, although some steps in the attachment process appear to disobey the requirements for such a mechanism.read more
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Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver.
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Spheroidal aggregate culture of rat liver cells: histotypic reorganization, biomatrix deposition, and maintenance of functional activities.
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TL;DR: It is concluded that hepatocytes cultured on matrigel, as opposed to the standard collagen, exhibit remarkably enhanced expression of many liver‐specific functions.
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Structure and function of heparan sulphate proteoglycans
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Formation of multicellular spheroids composed of adult rat hepatocytes in dishes with positively charged surfaces and under other nonadherent environments.
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TL;DR: Two distinct, mutually exclusive features of primary culture of adult hepatocytes apparently exist; monolayer culture with proliferative activity in an adherent environment and spheroid culture with poor proliferationative activity and high albumin-producing ability in a nonadherent environment.
References
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Role of collagenous matrices in the adhesion and growth of cells.
TL;DR: Current knowledge of the types of collagen and their distribution and biosynthesis is outlined, including fibronectin, which not only functions as an attachment protein, but also may play an important role in repair reactions.
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Fibronectins: multifunctional modular glycoproteins.
TL;DR: The current understanding of the structure and properties of fibronectins is reviewed to focus on overall concepts, recent developments and promising future research directions in this rapidly expanding field.
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Fibronectin: current concepts of its structure and functions.
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Isolation of a collagen-dependent cell attachment factor
Robert J. Klebe,Robert J. Klebe +1 more
TL;DR: It is demonstrated here that cell attachment to collagen is mediated by a high molecular weight protein present in serum, which is more important in cell attachment than the chemical composition of the plastic.
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Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions.
TL;DR: It is proposed that the method would be useful in preparation of fibronectin for studies on its biological activities, where it is important that the protein is obtained in a native state.