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Biomedical subjects

C Lapiere

Publications and source records attributed to C Lapiere.

6 recordsLinked to original sources

Serum and synovial fluid antibodies to collagen in rheumatic diseases: a review.

The literature on the occurrence and significance of antibodies to native or denatured collagen in rheumatic diseases is reviewed. Mainly type I and type II collagen have been investigated, both in serum and synovial fluid, with special reference to their possible role in rheumatoid arthritis. Brief results of the analysis of 40 synovial fluid samples, using an ELISA technique are included.

Arthritis, Rheumatoid

Synthesis of basement membrane components by differentiated thyroid cells.

Morphological studies indicate that basement membrane formation or maintenance can be achieved in cultures of thyroid cells. In the present investigation we have studied the biosynthesis of this extracellular matrix by differentiated porcine thyroid cells in culture. They were prepared by two procedures: (1) thyroid cells isolated by dispase digestion of the thyroid gland were maintained in serum-free medium on poly(L-lysine) coated dishes; (2) thyroid follicles released by collagenase treatment of the gland were isolated by differential filtration and cultured in suspension on agarose-coated dishes. In both cases, functional follicular-like structures were obtained as shown by their ability to organify Na125I and to respond to thyrotropin stimulation (250 microU/ml). After incubating the cells with radiolabeled proline or methionine, collagen synthesis was observed with the two types of culture, as shown by the formation of radioactive hydroxyproline and by the synthesis of peptides with electrophoretic properties identical to those of authentic collagen molecules and susceptible to collagenase. Besides variable amounts of type I and type III collagen-like peptides, significant proportions of labeled peptides migrated with type IV collagen chains and were precipitated by anti-type IV collagen antibody; thyrotropin had no significant effect either on the total collagen synthesis or on the relative amounts of the different collagen peptides. When thyroid cells were incubated with [35S]sulfate, a labeled glycosaminoglycan with chromatographic properties analogous to that of heparan sulfate could be obtained in both culture conditions; here again, no effect of thyrotropin was observed. The ability of differentiated porcine thyroid cells to synthesize basement membrane was suggested by their production of type IV collagen and heparan sulfate, two of its potential components. Thyrotropin, which drastically enhanced the functional property of the cells, did not seem to regulate this synthesis.

Animals

Collagen biosynthesis by cells in a tissue equivalent matrix in vitro.

Collagen biosynthesis by fibroblasts and subsequent processing and polymerization have been studied in conventional monolayer cultures and in a new model system in vitro in which cells organize into a tissue like structure. While the bulk of the newly made collagen becomes tightly and selectively bound to the matrix of the tissue equivalent model, in monolayer cultures most of the collagen passes into the culture medium. Collagen biosynthesis appears to be regulated differently in cells of the tissue equivalent model as compared with monolayered cells. In the former there is a 6- to 8-fold decrease in collagen output even though overall protein synthesis per unit of DNA is twice as great as in monolayered cells. Cells grown in tissue lattices exhibit much higher collagenolytic activity than cells in monolayer suggesting the model may also be of special use for studying collagen turnover and matrix remodeling.

Cell Line

The reconstitution of living skin.

A living-skin equivalent useful as a skin replacement and as a model system for basic studies has been fabricated and tested extensively. It consists of two components: (1) a dermal equivalent made up of fibroblasts in a collagen matrix that is contracted and modified by the resident cells, and (2) an epidermis that develops from keratinocytes "plated" on the dermal equivalent. A multilayered keratinizing epidermis with desmosomes, tonofilaments, and hemidesmosomes forms. Basement lamella formation occurs within 2 weeks in vitro when rat cells are used. With human cells, crypt or pseudofollicular morphogenesis is observed in vitro within 3 weeks after plating cells on the dermal equivalent. Autografts and isografts of rat-skin equivalents made with cultured cells from biopsies are rapidly vascularized, block wound contraction, and persist essentially for the lifespan of the host. Seven to 9 days after grafting, donor cells become activated biosynthetically and mitotically. By 1 year, the dermal population decreases to a normal level and the matrix has been extensively remodeled. The grafts remain free of hair and sebaceous glands. Grafts to rats have been in place for over 2 years. Now, allografts of dermal equivalents have been made across a major histocompatibility barrier and are not rejected. The persistence of cellular elements of the grafts is monitored by use of a genetic marker. Challenge of the allograft with a second skin-equivalent graft after 1 month does not result in rejection of the original graft or of the second skin-equivalent graft. We propose that allografts of tissue equivalents are tolerated because cells with class II antigens are selected against during in vitro cultivation and are excluded from the graft. Thus the fabrication of skin-equivalent tissues or of other equivalent tissues with parenchymal cells that do not bear class II antigens may render transplants of such tissues immunologically acceptable despite the presence of allogeneic cells. The capacity to graft across major histocompatibility barriers using living tissue equivalents may have important clinical significance.

Animals