Scanning Electron Microscopy


Collagenous matrices, used as cell culture substrata, can be prepared from different collagen types in a variety of forms using a range of polymerization procedures. Type I collagen has been most frequently used either as dried collagen films or hydrated collagen gels. Sheets of isolated bladder urothelium, when plated onto such matrices prepared from type I collagen by different polymerization methods (eg. air-drying; NaOH; NaCl; NH3; or NH3 followed by glutaraldehyde crosslinking) demonstrate the capability of urothelial cells to attach to a variety of differently prepared matrices irrespective of polymerization procedure. In contrast, both cell proliferation and maintenance of the urothelium are markedly influenced by the polymerized form of the collagen matrix.

Comparative ultrastructural (scanning and transmission electron microscopy) analysis of these matrices demonstrates dissimilarities in their physical organization. The level of filamentous, fibrillar or fibrous reaggregation of solubilized collagen molecules varies in relation to the polymerization procedure used viz, a) air dried matrices form a dense meshwork of many forms of collagen fibrils and associated filaments with an irregular surface array of coarser collagen fibres; b) matrices prepared by NaOH, NaCl and NH3 polymerization present no major differences and form a felt of interlocking collagen fibres with discrete filamentous networks associated with these fibres; and c) matrices polymerized by NH3 and crosslinked with glutaraldehyde form a dense meshwork of filaments with a more occasional distribution of fibrils associated with filaments or dense "amorphous" aggregates.

The level of supramolecular reassemblage of solubilized collagen may be, therefore, a significant factor in determining urothelial cell growth and differentiation on collagen matrices.

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