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P Van Gansen

Publications and source records attributed to P Van Gansen.

17 recordsLinked to original sources
PubMed1992

Transdifferentiation of macrophages into fibroblasts as a result of Schistosoma mansoni infection.

The possibility of transdifferentiation of macrophages into fibroblasts which could be at the origin of fibrotic tissue in schistosome-infected mice was studied using immunocytochemical techniques. Macrophage cell samples extracted from the peritoneal cavity of schistosome-infected mice were fractionated on a Percoll gradient. The cultures were purified by treatment with a trypsin solution to eliminate any fibroblasts possibly collected along with the macrophages. Immunocytochemical methods were then used to characterize the cells at different points in time. The fibroblastic property of the morphologically transformed cells was confirmed by their positive labeling with the anti-procollagen antibody. However, these cells still possessed the mac-1 and mac-2 antigens which characterize the monomacrophage line.

Animals↗
PubMed1992

A study of peritoneal cells from healthy and Schistosoma mansoni-infected mice with special reference to myofibroblasts arising in culture.

Adherent, trypsin-resistant, peritoneal cells from mice with chronic schistosomiasis mansoni, and from control mice, were cultivated in vitro up to 20 days. Fibroblasts regularly appeared, about 6 days after seeding, in cultures of the manyfold more numerous cells from infected mice, concomitantly with a dramatic increase, detected by autoradiography, in the percentage of DNA-replicating cells of the monocyte-macrophage lineage. Peritoneal cells from healthy and from infected mice were fractionated on discontinuous Percoll gradients. Eight cell subsets were harvested in both cases, quantitated, and studied by electron microscopy. Two fractions (2 and 3: 1.041 < densities < 1.060 g/ml) from infected mice were greatly enriched in monoblasts and promonocytes. The cells of the different subsets were seeded separately, trypsin-treated and cultivated in vitro. Cultures of cell fractions 2 and 3 from infected mice contained the majority of the DNA-synthesizing cells and gave regularly rise to fibroblasts. Cultures of the different fractions were used for sequential morphological observations (2-11 days) at the electron microscope level. Early cultures were also used for the ultrastructural detection of the Mac-1 (CD 18/CD 11b) surface antigen by gold immunocytochemistry. A few fibroblasts were rarely observed in cultures of fractions 2 and 3 from control mice, while cells with ultrastructural features of myofibroblasts were regularly observed in cultures of the same fractions harvested from mice with chronic schistosomiasis. Fractions 2 and 3 from infected mice contained a large number of Mac-1 positive monoblasts. The correlations between the presence of monoblasts, DNA replication in cells of the monocyte-macrophage lineage and the appearance of myofibroblasts in cultures of the same fractions derived from infected mice are discussed.

Animals↗
PubMed1991

Eosinophil granulocytopoiesis in hepatic periovular granulomas during the chronic phase of experimental murine Schistosomiasis mansoni.

We have observed in hepatic periovular granulomas of C3H mice infected with Schistosoma mansoni, in the chronic phase of the disease (12-19 weeks of infection), groups of early precursors and immature eosinophil granulocytes corresponding, at the ultrastructural level, to promyelocytes and myelocytes. Mitosis was also seen in eosinophil myelocytes. These eosinophil myeloid foci were observed in close contact with macrophages and epithelioid cells, and they were surrounded by an extracellular matrix, rich in collagen fibres. These morphological observations give support to the concept of a peripheral proliferation of eosinophils in chronic schistosomiasis, mediated by a factor secreted by macrophages present in granulomas.

Animals↗
PubMed1989

Schistosomiasis and in vitro transdifferentiation of murine peritoneal macrophages into fibroblastic cells.

We developed a method for avoiding contamination by fibroblasts when cultures of peritoneal cells are initiated. Macrophages were identified by immunogold detection [light microscope, transmission (TEM) and scanning (SEM) electron microscopes] of membrane antigens (Mac-1+, Thy-1,2-), non-specific esterase activity and ultrastructural features (TEM). As compared with controls, the yield of peritoneal macrophages was 2- and 12-fold higher, respectively, in acutely and chronically infected mice. In all, 30 "chronic", 18 "acute" and 18 control cultures were followed up. At a given cell-density seeding, the decline of control, "acute" and "chronic" cultures starts at about day 10, 15, and 27, respectively. In "chronic" cultures only, fibroblast-like cells appear from day 6 onwards; their number increases with time. Cells showing characters intermediary between macrophages and fibroblasts were observed. We suggest that fibroblast-like cells result from the in vitro transdifferentiation of a limited number of in vivo committed macrophages.

Animals↗
PubMed1988

Potential and limitations of cultivated fibroblasts in the study of senescence in animals. A review on the murine skin fibroblasts system.

Senescence is the last period of the life span, leading to death. It happens in all animals, with the exception of a few didermic species (Hydras) having a stock of embryonic cells and being immortal. The causes of animal senescence are badly known. They depend both on genetic characters (maximum life span of a species) and on medium factors (mean expectation of life of the animals of a species). Animal senescence could depend on cell aging: (1) by senescence and death of the differentiated cells, (2) by modified proliferation of the stem cells of differentiated tissues, (3) by alterations in the extracellular matrices, (4) by interactions between factors (1) (2) and (3) in each tissue, and (5) by interactions between the several tissues of an organism. This complexity badly impedes the experimental study of animal senescence. Normal mammal cells are aging when they are cultivated (in vitro aging). Present literature upon in vitro aging of cultivated human fibroblasts consists essentially of papers devoted to proliferation and differentiation characteristics and not to cell senescence. Murine skin fibroblasts have been studied in our laboratory, using different systems: (1) primary cultures isolated from peeled skins of mouse embryos, (2) mouse derms analysed in the animals, (3) cultivated explants of skins, (4) serial sub-cultures of fibroblasts isolated from these explants, (5) cells cultivated comparably on plane substrates (glass, plastic, collagen films) and on three-dimensional matrices (collagen fibres). In primary cultures (system 1) all the cell generations have been analysed, including the last one until death of the culture. We have shown that many characters are varying with cell generation. All the observed variations were: progressive, non-linear and correlated (intracellular feedbacks). We come to the conclusion that the main effects of cell mitotic age are (1) to depress the plasticity of the chromatin, (2) to change the organization of the cytoplasmic filaments, (3) to change the organization of the extracellular matrix. The collagen fibres are also acting upon nucleus and filaments either in the animals or in the cultures. The phenotype of a fibroblastic cell is thus both age- and environment-dependent. Overall data on in vitro cell aging point to the hypothesis that senescent cells are phenotypic variants and not mutant cells. Aging cell cultures are remarkably useful to the studies on cell proliferation decrease and cell cycle lengthening shown by the stem cells in animal tissues. We propose the hypothesis that the fibroblasts of the vertebrates would be homologous to the pluripotent mesenchyme cells of their embryos.

Actin Cytoskeleton↗
PubMed1987

[Possibilities and limitations of fibroblast cultures in the study of animal aging].

INTRODUCTION. Aging--the effect of time--occurs in every living organism. Senescence is the last period of the lifespan, leading to death. It happens in all animals, with the exception of a few didermic species (Hydras) having a stock of embryonic cells and being immortal. The causes of animal senescence are badly known. They depend both on genetic characters (maximal lifespan of a species) and on medium factors (mean expectation of life of the animals of a species). Animal senescence could depend on cell aging: 1) by senescence and death of the differentiated cells, 2) by modified proliferation and differentiation of the stem cells of differentiated tissues, 3) by alterations in the extracellular matrices, 4) by interactions between factors 1) 2) and 3) in each tissue, 5) by interactions between the several tissues of an organism. This complexity badly impedes the experimental study of animal senescence. Normal mammal cells are aging when they are cultivated (in vitro ageing): their phenotype varies and depends on the cell generation (in vitro differentiation); the last cell-generation doesn't divide anymore and declines until death of the culture (in vitro senescence). Analysis of these artificial but well controlled systems allows an experimental approach of the proliferation, differentiation, senescence and death of the cells and of the extracellular matrix functions. Present literature upon in vitro aging of cultivated human cells is essentially made of papers where proliferation and differentiation characteristics are compared between early ("young") and late ("old") cell-generations of the cultures. FIBROBLASTIC CELLS OF THE MOUSE SKIN. This cell type has been studied in our laboratory, using different systems: 1) Primary cultures isolated from peeled skins of 19 day old mouse embryos, 2) Mouse dermis analyzed in the animals, 3) Cultivated explants of skins, 4) Serial sub-cultures of fibroblasts isolated from these explants, 5) Cells cultivated comparably on plane substrates (glass, plastic, collagen films) and on tridimensional matrices (collagen fibres). Systems 2), 3), 4) and 5) have been obtained either from 19 day old embryos or from 6 groups of animals of different ages (from 1/2 till 25 month). In primary cultures (system 1) all the cell generations have been analyzed, including the last one until death of the culture. We have shown that many characters are varying with cell-generation: cell form and cell mass, rate of DNA replication and cell division, rate of RNA transcription, nature of the accumulated and of the synthetized proteins, organization of the cytoskeletal elements, organization of the extracellular matrix, type of cell death.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗
PubMed1986

Fibronectin localization and endocytosis in early and late mouse embryonic fibroblasts in primary culture: a study by light and electron microscopic immunocytochemistry.

In spreading fibroblasts, strong endocytosis of exogenous fluorescent fibronectin (FN) was observed from the beginning of their attachment to the substratum. In early fibroblasts, the internalized FN was localized both in the peripheral ruffles and in the perinuclear cytoplasm; in late fibroblasts, whose spreading was slower, FN uptake was not detected in the ruffles. In growing cultures, supracellular FN fibres, detected by direct fluorescence microscopy or by the indirect peroxidase-anti-peroxidase (PAP) complex technique, were scarce on early cells, but very numerous on the upper face and on the filopodia of late cells. At the ultrastructural level, FN, localized with the immuno-gold staining method, was found associated with fibres of the extracellular matrix and the upper face of the cells. FN was endocytosed via smooth vesicles and we suggest that the internalization process is slower in the late cells. In confluent early cultures, an extended network of pericellular FN was observed as usual. The pericellular FN of late grouped cells was present as a few coarse fibres connected with some of the cell surface threads.

Animals↗
PubMed1985

Actin content and organization of microfilaments in primary cultures of mouse embryonic fibroblasts (in vitro ageing).

Actin distribution in serially passaged embryonic mouse fibroblasts has been visualized by the anti-actin-PAP method; the organization of the microfilaments has been observed by electron microscopy (SEM and TEM). Four successive actin patterns have been identified: early (few well-organized bundles of microfilaments), middle-aged (many well-organized bundles and patches around the nucleus), late (numerous ill-organized filamentous structures and diffuse perinuclear-actin) and "senescent" (heavy packs of short microfilaments around the nucleus). All the observed actin-positive filaments were disrupted by cytochalasin B treatment. The cytoplasmic actin complex was cell-age and not cell-size-dependent; it behaved differently from the cytoplasmic microtubular complex to serially subcultivated fibroblasts. Measurements of the cell-protein content (Lowry's method) and SDS-polyacrylamide gel electrophoresis (Laemmli's method) have been performed in the successive population doubling levels (PDL) of the primary cultures. Triton-insoluble actin increased in parallel with total protein and reached about 4% of the total proteins in all the PDLs. Triton-soluble actin also increase at the beginning of the middle-aged period (generally 6 PDL) and another in declining cultures (generally 10 PDL). Total actin amounted to about 8% of the total proteins in early fibroblasts, to about 16% at the beginning of the middle-aged period and to about 20% in the declining terminal cultures. Taking into account all the known characteristics of subcultivated primary cultures, we tentatively consider the evolution of the fibroblasts as an in vitro differentiation followed by true in vitro senescence in the declining cultures. Regarding the cytoplasmic actin-complex, senescence would be characterized by a sharp increase in soluble actin, an unbalanced ratio between soluble and insoluble actin and an impairment of the ability of the microfilaments to form well-organized bundles.

Actin Cytoskeleton↗
PubMed1979

[Cell aging in vitro].

The phenomenon of in vitro cell ageing has been considered in terms of limited multiplication and defined life span of cultured diploid cell strains and lines in vitro and in situ. Theories on cell ageing are discussed. Original recent evidence is provided concerning mouse embryo fibroblast ageing in vitro. Phenotypes of early, middle and late passage cells, observed with transmission and scanning electron microscopes, are described. Using light microscope autoradiography, proliferative capacity of these cells has been examined that involves replicative index, cell cycle durations, and 3H-actinomycin binding index. Transcription peculiarities of cultured cells of different age are described using such parameters as activities of RNA polymerases and of RNA synthesis estimated biochemically in isolated nuclei, and with the help of light autoradiography in living cells.

Aging↗
PubMed1978

Cytochemical study of human lymphocytes stimulated by PHA in function of donor age.

The 3H-AM binding reflects the structural changes involved in the cellular differentiation. This parameter was studied during blastic transformation of human lymphocytes, in relation to the age of the donor. Although they are individual variations, the 3H-AM binding is higher in the young group than in the aged subjects, as well as the blast transformation score. These results indicate that the weak lymphocyte response to the mitogen lectine (PHA) stimulation could be related to some age-induced structural alterations of the chromatin, resulting in an irreversible blockage in G1, at least in some of the T lymphocytes.

Adult↗