Cultured corneal fibroblasts as a model system for the demonstration of drug-induced mucopolysaccharidosis

Burmester, J.; Handrock, K.; Lüllmann‐Rauch, Renate

doi:10.1007/bf01972989

Cited by 18 publications

(6 citation statements)

References 22 publications

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“…The purity of control astroglial monolayers was additionally screened with antibodies to detect microglia, oligodendrocytes, fibroblasts and endothelial cells. Fibroblasts were prepared according to standard procedures (Burmester et al 1990) from corneal explants from young postnatal rats.…”

Section: Cell Preparation and Culture Conditionsmentioning

confidence: 99%

Ramification of microglia, monocytes and macrophages in vitro: influences of various epithelial and mesenchymal cells and their conditioned media *

Wilms

Hartmann

Sievers

1997

Cell and Tissue Research

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Microglial cells are able to switch between an "active" amoeboid and a ramified "resting" morphology during development and after experiencing lesions. We have previously shown that in vitro microglial morphology is controlled by their cellular environment, i. e. cells become ramified in astrocyte coculture but amoeboid on monolayers of fibroblasts. In the present study we have extended the analysis of the control of macrophage morphology by maintaining macrophages of different origins in coculture with different epithelial or mesenchymal cells and their conditioned media. Microglia, monocytes and spleen macrophages seeded onto monolayers of astrocytes, kidney epithelia or hepatoma cells developed the ramified morphology but remained amoeboid in fibroblast coculture. Ramification was also induced by media conditioned by these cells as well as by phorbolic esters, i.e. activators of protein kinase C. In double coculture assays, even small numbers of fibroblasts were able to override the "epithelial" influence. Likewise, microglia remained amoeboid, when incubated on several constituents of the extracellular matrix. These results indicate that macrophage ramification is an active process initiated by diffusible factors secreted by various epithelial cells, possibly acting upon a protein-kinase-C-related receptor. We interprete the modification of macrophage morphology as a functional adaptation to the surrounding type of tissue that is enforced by its constituent cells. Thus, the specific morphologies of microglia, hepatic von Kupffer's cells or peritubular kidney macrophages could be explained by similar epithelium-macrophage interaction.

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Section: Cell Preparation and Culture Conditionsmentioning

confidence: 99%

Ramification of microglia, monocytes and macrophages in vitro: influences of various epithelial and mesenchymal cells and their conditioned media *

Wilms

Hartmann

Sievers

1997

Cell and Tissue Research

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“…Monolayers of fibroblasts were prepared according to standard procedures (Burmester et al, 1990) from either explants of corneae or cell suspensions of dissociated meninges, both from young postnatal rats.…”

Section: Preparation Of Fibroblast Monolayersmentioning

confidence: 99%

Blood monocytes and spleen macrophages differentiate into microglia‐like cells on monolayers of astrocytes: Morphology

Sievers

Parwaresch

Wottge

1994

Glia

133

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Several morphological and functional properties of microglial cells, the resident immunoeffector cells of the central nervous system (CNS), differ from those of monocytes/macrophages in other tissues. Microglia are assumed to derive from myelonocytic lineage, possibly as a distinct subpopulation that diverges from a common cell line early in ontogeny, invades the CNS, proliferates, and differentiates into ameboid and then ramified microglia. We tested the hypothesis that some morphological and functional properties of microglia are induced in myelomonocytic cells by nervous tissue, specifically astrocytes. In the present in vitro studies we compared the differentiation of microglia, blood monocytes, and spleen macrophages on acellular substrates and on monolayers of astrocytes and fibroblasts. On acellular substrates, microglial cells at first acquire an ameboid morphology; later they show a few short, unbranched processes. On monolayers of pure astrocytes, microglial cells at first also differentiate into ameboid cells, but after 5 to 7 days they start to develop processes with large lamellopodial tips. These lengthen and branch continuously during the next 2 weeks in vitro, demarcating a round to oval territory around the small ellipsoid cell body. By contrast, on monolayers of fibroblasts the microglial cells develop an ameboid morphology, but do not grow the typical long branched processes of the ramified form. Blood monocytes and spleen macrophages behave indistinguishably from microglia both on acellular and cellular substrates, i.e., on astroglia they develop the ramified form, while on fibroblasts they retain the ameboid shape. When microglia, macrophages, or monocytes are cultured on coverslips on top of astrocytic monolayers, i.e., physically separated from the astroglia, but exposed to the medium conditioned by astrocytes, a significant proportion of them also develop the ramified shape. These findings indicate that the ramified shape of microglia is induced by astrocytes. Since this morphology can also be induced in blood monocytes and macrophages, we take this to be further evidence for the proposition that microglial cells are derived from the myelomonocytic lineage, and, moreover, that properties of resident macrophages are largely determined by tissue components of their host organ.

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“…Exposure to the drug usually lasted 72 h, with no change of the culture medium. This time period was previously found to be most appropriate for inducing marked GAG storage with tilorone and its bisbasic analogues [7,14].…”

Section: Cell Cultures and Experimental Conditionsmentioning

confidence: 98%

“…MT was synthesized by the late Professor R. Haller (Department of Pharmaceutical Chemistry, University of Kiel) according to a published method [11]. The materials and media for cell culture work were the same as used previously [14]. Na # $&SO % (specific radioactivity 10-100 mCi\mmol) was purchased from Amersham-Buchler (Braunschweig, Germany).…”

Section: Experimental Materialsmentioning

confidence: 99%

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Tilorone-induced lysosomal lesions: the bisbasic character of the drug is essential for its high potency to cause storage of sulphated glycosaminoglycans

Fischer

Hein

Lüllmann‐Rauch

et al. 1996

Biochemical Journal

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The immunomodulatory agent tilorone -2,7-bis-[2-(diethyl-amino)ethoxy]fluoren-9-one- and congeners are potent inducers of lysosomal storage of sulphated glycosaminoglycans (GAGs) in animals and cultured fibroblasts of animals and man. All potent inducers of GAG storage hitherto described are bisbasic polycyclic aromatic compounds. They are accumulated in lysosomes and disturb the degradation of GAGs, mainly dermatan sulphate. It has been proposed that the drugs cross-link the polyanionic GAG chains giving rise to undergradable drug-GAG complexes. This hypothesis implies that the bisbasic character of the drug molecules is essential for the side effect in question. In the present study, this was tested by comparing tilorone and its monobasic derivative (MT) with respect to (i) induction of GAG storage in cultured bovine corneal fibroblasts and (ii) physicochemical interactions with GAGs in vitro. The intralysosomal concentration of MT achieved after 1-3 days was of the same order of magnitude as previously shown for tilorone. Nevertheless, under conditions that did not enhance the secretion of a lysosomal enzyme (beta-hexosaminidase, EC 3.2.1.52), the ability of MT to cause storage of [35S]GAGs was significantly lower than that of tilorone. Morphological observations showed that MT was much more potent in causing lysosomal storage of polar lipids than of GAGs. CD spectroscopy with tilorone revealed that the presence of GAGs caused the primarily achiral drug molecules to display CD. This suggested a helical orientation of the tilorone molecules within GAG-drug complexes, and short intermolecular distances which allowed electronic coupling of the aromatic ring systems of adjacent drug molecules. In contrast, MT failed to display any induced optical activity, indicating the absence of highly ordered GAG-drug complexes. In conclusion, the present results show that the substitution of the planar aromatic ring system with two basic side chains is essential for the high potency of tilorone in inducing lysosomal GAG storage. This is paralleled by, and presumably causally related to, strong physicochemical interactions with GAGs.

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Cultured corneal fibroblasts as a model system for the demonstration of drug-induced mucopolysaccharidosis

Cited by 18 publications

References 22 publications

Ramification of microglia, monocytes and macrophages in vitro: influences of various epithelial and mesenchymal cells and their conditioned media *

Ramification of microglia, monocytes and macrophages in vitro: influences of various epithelial and mesenchymal cells and their conditioned media *

Blood monocytes and spleen macrophages differentiate into microglia‐like cells on monolayers of astrocytes: Morphology

Tilorone-induced lysosomal lesions: the bisbasic character of the drug is essential for its high potency to cause storage of sulphated glycosaminoglycans

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