Cell Culture Models: Reliable Tools in Pharmacotoxicology?

Fusenig, N. E.

doi:10.1007/978-3-662-03011-0_1

Cited by 7 publications

(5 citation statements)

References 6 publications

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“…Likewise, organotypic cocultures have proved well-suited systems to test the effects of differentiation modulators, growth as well as nutritive factors (Choi and Fuchs, 1990;Boyce and Williams, 1993;Ponec et al, 1997a). Besides their role in basic skin biology research, cultured skin equivalents are valuable tools for pharmacologic and toxicologic in vitro testing as an alternative to animal models (Gay et al, 1992;Fusenig, 1994b; and for therapeutic use as skin substitutes (e.g., wound coverage, gene therapy) (reviewed by Boyce, 1996). In all these fields of application, standardization of the in vitro system is required.…”

mentioning

confidence: 99%

Organotypic Keratinocyte Cocultures in Defined Medium with Regular Epidermal Morphogenesis and Differentiation

Stark

Baur

Breitkreutz

et al. 1999

Journal of Investigative Dermatology

200

221

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Skin equivalents formed by keratinocytes cocultured with fibroblasts embedded in collagen lattices represent promising tools for mechanistic studies of skin physiology, for pharmacotoxicologic testing, and for the use as skin substitutes in wound treatment. Such cultures would be superior in defined media to avoid interference with components of serum or tissue extracts. Here we demonstrate that a defined medium (supplemented keratinocyte defined medium) supports epidermal morphogenesis in organotypic cocultures equally well as serum-containing medium (mixture of Ham's F12 and Dulbecco's modified Eagle's medium), as documented by hallmarks of the epidermal phenotype studied by immunofluorescence and electron microscopy. In both cases regularly structured, orthokeratinized epithelia evolved with similar kinetics. Morphology in mixture of Ham's F12 and Dulbecco's modified Eagle's medium was slightly hyperplastic, and keratins 1 and 10 synthesis less co-ordinated than in supplemented keratinocyte defined medium, but a consistently inverted sequence of expression of keratins 1 and 10 was found in either medium. The late differentiation markers filaggrin, involucrin, keratin 2e, and transglutaminase 1 corresponded in their typical distribution in upper suprabasal layers. Keratin 16 persisted under both conditions indicating the activated epidermal state. Keratinocyte proliferation was comparable in both media, whereas fibroblast multiplication and proliferation was delayed and reduced in supplemented keratinocyte defined medium. In both media, ultrastructural features of epidermal differentiation as well as reconstitution of a basement membrane occurred similarly. Immature lamellar bodies and cytoplasmatic vacuoles, however, indicated an impaired lipid metabolism in supplemented keratinocyte defined medium. Nevertheless, these defined organotypic cocultures provide a suitable basis for in vitro skin models to study molecular mechanisms of tissue homeostasis and for use in pharmacotoxicologic testing.

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mentioning

confidence: 99%

Organotypic Keratinocyte Cocultures in Defined Medium with Regular Epidermal Morphogenesis and Differentiation

Stark

Baur

Breitkreutz

et al. 1999

Journal of Investigative Dermatology

200

221

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“…Dermal equivalents were prepared with collagen type I extracted from rat tail tendons and dermal fibroblasts (passage 5–9) at final concentrations of 3·4 mg mL −1 collagen and 1·7 × 10 5 fibroblasts mL −1 /1× Hanks/10% FBS. A 3‐mL aliquot of this mixture was poured into BD Falcon cell culture inserts (Becton Dickinson Co., Franklin Lakes, NJ, U.S.A.) in six‐well BD Falcon companion plates and allowed to jellify for 1 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

A mouse organotypic tissue culture model for autosomal recessive congenital ichthyosis

et al. 2014

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The mouse skin equivalents faithfully recapitulate the 12R-LOX-deficient phenotype observed in vivo, classifying them as appropriate in vitro models to study molecular mechanisms involved in the development of ARCI and to evaluate novel therapeutic agents. In contrast to existing human three-dimensional skin models, the generation of these murine models is not constrained by a limited supply of material and does not depend on in vitro expansion and/or genetic manipulations that could result in inadvertent genotypic and phenotypic alterations.

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“…The use of such genetically modified cell lines, for example, carrying promoters for immediate response genes, coupled with reporter genes which give rise to products that are easily and quantitatively measurable, will considerably improve our in vitro toxicology technology. When these genetically modified cell lines can be cultured under optimal organotypic culture conditions (enabling them to form a normal functioning epidermis), they will become a very promising and attractive in vitro test system (46).…”

Section: Immortalised Keratinocyte Cell Linesmentioning

confidence: 99%

The Use of Human Keratinocytes and Human Skin Models for Predicting Skin Irritation

et al. 1999

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Cell Culture Models: Reliable Tools in Pharmacotoxicology?

Cited by 7 publications

References 6 publications

Organotypic Keratinocyte Cocultures in Defined Medium with Regular Epidermal Morphogenesis and Differentiation

Organotypic Keratinocyte Cocultures in Defined Medium with Regular Epidermal Morphogenesis and Differentiation

A mouse organotypic tissue culture model for autosomal recessive congenital ichthyosis

The Use of Human Keratinocytes and Human Skin Models for Predicting Skin Irritation

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