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Biomedical subjects

M Prunieras

Publications and source records attributed to M Prunieras.

At least 37 records · Page 2Linked to original sources
PubMed1984

Human Langerhans cells in epidermal cell culture, in vitro skin explants and skin grafts onto "nude" mice.

In order to find a model system which best preserves human Langerhans cells (LC) outside of the human body, three possibilities were examined: epidermal cell culture, skin explants, and skin grafts onto "nude" mice. Using OKT-6 and anti-HLA-DR monoclonal antibodies, we quantified LC in epidermal sheets or epidermal cell cultures. All observations were carried out over a period of 4 weeks. We found that under epidermal cell culture conditions, LC rapidly disappeared, to the extent that after 10 days only rare HLA-DR-positive cells could be observed. In contrast, in the presence of intact dermis (explants and grafts), 60%-80% of the original number of LC, morphologically unchanged, dendritic and OKT-6 and HLA-DR-positive, were seen. These findings suggest that human LC are either a long-lived cell population or else can proliferate locally. The systems studied may be a useful tool for future investigation of LC function.

Animals↗
PubMed1984

A keratin of fetal skin is reexpressed in human keratinocytes transformed by SV40 virus or treated with the tumor promoter TPA.

SV40-transformation as well as treatment with tumor promoters produce alterations in morphology, differentiation and keratinization of human keratinocytes. Two cell lines of SV40-transformed keratinocytes and primary cultures of keratinocytes treated with the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate (TPA) were found to contain an additional protein of 52.5 kD molecular weight (MW). This protein was identified by its reactivity with the monoclonal antibody TROMA-I as being keratin no. 8, a keratin normally present only in simple epithelia. Since this keratin is present in fetal epidermis but disappears gradually when fetal skin becomes multilayered after week 13 of development (Moll et al., Differentiation 23 (1982) 170. [23]), it suggests that SV40 virus and TPA are able to induce in human keratinocytes the reexpression of fetal characters.

Cell Line↗
PubMed1984

An improved method for the establishment of cell lines from primary skin melanomas.

Fourteen explants of primary skin melanomas and six explants of lymph node metastases were used to establish melanocyte cell lines, and in all twenty experiments the melanocytes became overgrown by fibroblasts. By contrast, eight melanocyte cell lines without fibroblast contamination were established by mechanical cell separation followed by either direct plating in microwells (five out of thirteen primary tumours and three out of four lymph node metastases) or plating of cells from colonies grown in soft agar (three out of five cases). It was also observed that cell crowding and partial (as opposed to total) replacement of the culture medium during the early steps of culture favoured the growth of tumour cells. This observation suggests that melanoma cells are subjected to an autocrine type of growth regulation.

Cell Line↗
PubMed1983

Antimelanoma hybridoma antibodies against partially purified melanoma antigen.

We have immunised BALB/c mice with a melanoma antigen obtained after papain solubilisation of the membranes of a metastatic melanoma tumour and fused the immune spleen cells to the mouse myeloma line P3-NS1/1-Ag4.1. The produced hybridoma antibodies (Mel-PV antibodies) recognised the initial melanoma antigen in haemagglutination, but did not react with any of the HLA phenotypes tested by cytotoxicity on a panel of B lymphocytes with known HLA-A and B phenotypes. We rosetted red blood cells coated with protein A with dispersed cells from fresh melanoma tumours, and a high degree of specificity for human malignant melanocytes was observed. Purified Mel-PV antibodies were also tested by indirect immunofluorescence and found to be oriented towards cytoplasmic components of malignant melanoma cells. These results indicate that the use of melanoma antigens for preparing monoclonal antibodies maintained a satisfactory degree of specificity and may be an adequate starting point for defining common and specific antigenic determinants on human melanoma.

Animals↗
PubMed1982

Effect of phorbol esters on guniea pig skin in vivo.

When topically applied to guniea pig ear skin the tumor promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced inflammation and epidermal hyperproliferation which could be inhibited by indomethacin. This inhibition could be reversed both by prostaglandins E and F. Five minutes after TPA treatment an increase in the level of prostaglandin E but not of prostaglandin F was observed in the epidermis. The non-promoting phorbol ester 4-O-methyl-TPA also stimulated epidermal cell proliferation but this stimulation was not inhibited by indomethacin. The above results are in agreement with those already reported in the mouse system with these two compounds. Ornithine decarboxylase (ODC) activity has been evaluated in the epidermis of guniea pig ear after topical application of 20 nmol of TPA. No increase was noted. This is in contrast with the well documented activation of ODC in mouse skin treated with TPA. Since TPA acts as a promoter in the mouse whereas both croton oil and TPA have no promoting action in the guinea pig, the above result supports the view that ODC activationis related to promotion, and provides a possible explanation for the resistance of this animal species to promotion. This resistance is further documented by the fact that no "dark cells" were found in guinea pig ear skin.

Animals↗
PubMed1981

Skin explant cultures: expression of cytoplasmic differentiation antigens in outgrowth cells.

In vivo, keratinocyte cytoplasmic antigens linked to keratinization have been traced with 2 distinct types of antibodies found in human sera. One type of antibody is specific for the basal cell compartment of keratinocytes while the other one is specific for upper keratinocytes. Using these markers, we followed a well-defined, in vitro human keratinocyte culture system that produces a keratinizing epithelial cell outgrowth juxtaposed to dead dermal substrate for the in vitro expression of keratinocyte cytoplasmic antigens. These antigens were found to be expressed in vitro independently from dermal influences. Their chronology, localization and topography in culture matched the in vivo situation.

Antigens↗
PubMed1980

Bullous pemphigoid antigen synthesized in vitro by human epidermal cells.

Cultured living human epidermal cells migrating on the surface of irradiated non-viable pig dermis, were found to produce bullous pemphigoid antigen at the epidermis-pig dermal junction between days 10 and 50 in culture. In certain series, the antigen was detected within the cytoplasm of the basal cells as well. These findings suggest that the bullous pemphigoid antigen is synthesized by epidermal cells.

Animals↗
PubMed1979

Recent advances in epidermal cell cultures.

Among the many skin culture systems, three have been selected in this short review because of their specific potentials in dermatological research. H. Green cultures newborn human forsekin keratinocytes on a mouse 3T3 feeder layer. Keratinocytes grow and keratinize. The feeder cells release factor(a) which allows serial propagation of keratinocytes to be achieved. The cell yield is further increased by adding epidermal grohth factor. This system has already proved to be a potent tool for the study of keratinization at the molecular level. A. Freeman has described a system in which explants of adult human skin are cultured on the dermal aspect of dead split-thickness pig skin. Keratinocytes can be passaged several times. Their differentiation is remarkable: it includes the production of keratohyaline, membrane coating granules, pemphigus as well as pemphigoid antigens. This system is interesting in the study of epidermal morphogenesis and may be applicable to the treatment of burns. The culture of epidermal cells from adult guinea pig ear in comparison with that of dermal fibroblasts is being used to study the specificity of action of pharmacological compounds on growth and keratinization of epidermal cells. Furthermore, the isolation (and culture) of pure populations of basal cells appears as a promising approach to the study of the mechanisms which moderate epidermal cell proliferation.

Animals↗
PubMed1979

Microflowfluorometric evaluation of homeostasis in cultured epidermal cells.

In isolating and culturing in vitro populations of basal cells from adult guinea pig skin, it has been possible to show that these cells are sensitive to both G1 and G2 inhibitions. Only a small fraction (10 percent or less) of the G1 blocked cell population would be governed by G1 inhibitory messages released by suprabasal, maturing keratinocytes. As regards the G2 block in vitro experiments confirm that basal cells produce a G2 blocker to which about 9 per cent or less are susceptible. In conclusion basal cells in culture are sensitive to homeostatic regulation as in vivo.

Animals↗