Episkin, a reconstituted human epidermis for assessing in vitro the irritancy of topically applied compounds

Roguet, R.; Cohen, C.; Dossou, K.G.; Rougier, A.

doi:10.1016/0887-2333(94)90195-3

Cited by 70 publications

(36 citation statements)

References 20 publications

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“…Upon receipt of the EPISKIN kit (on day 13 of culture), the culture inserts ( fig. 1 ) were removed from their nutrient gel and transferred under aseptic conditions into a sterile culture COSTAR plate of 12 wells (Corning Inc., Corning, N.Y., USA) containing 1.5 ml of a maintenance medium (EPISKIN SNC) [15] per well. The samples were then incubated at 37 ° C with 5% CO 2 and at 98% relative humidity.…”

Section: Episkinmentioning

confidence: 99%

Improvement of the Experimental Setup for Skin Absorption Screening Studies with Reconstructed Skin EPISKIN®

Grégoire

Patouillet²,

Noé³

et al. 2008

Skin Pharmacol Physiol

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Percutaneous penetration studies are usually performed in human skin samples set up in a Franz® cell device. The ability to perform these studies may depend on the availability of skin samples. Reconstructed skin models are an interesting alternative to overcome such limitations but are less easily mounted in diffusion cell devices. Previous data showed that EPISKIN® was a highly performing model to carry out such studies. However, the setup in a PermeGear® cell device is time consuming and therefore unsuitable for screening purposes. Another approach could be using EPISKIN in its cell culture insert. The aim of this study was to compare cutaneous penetration of chemicals applied to EPISKIN samples in a PermeGear cell versus in their own insert. Eight chemicals having widely different chemical structures and penetration potentials were studied. Six test chemicals showed a similar penetration level in both devices. Using the PermeGear cell device, the penetration level was overestimated for the other 2 tested chemicals. The results demonstrated that percutaneous studies with EPISKIN samples could be easily performed using the insert setup. The EPISKIN model has been greatly improved in the recent years and it is now possible to develop screening tests for the evaluation of skin penetration with a higher reliability.

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Section: Episkinmentioning

confidence: 99%

Improvement of the Experimental Setup for Skin Absorption Screening Studies with Reconstructed Skin EPISKIN®

Grégoire

Patouillet²,

Noé³

et al. 2008

Skin Pharmacol Physiol

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“…Upon receipt of Episkin cultured for 20 days, skin samples were removed from their nutritive gel and transferred under aseptic conditions into sterile 12-well culture dishes containing 2 ml of maintenance medium per well (Episkin SNC) [16]. Then the cultures were incubated at 37°C, 5% CO 2 , and 98% humidity until use the next day.…”

Section: Skin Sources and Storage Conditions Epiderm Epi-606 (Mattek mentioning

confidence: 99%

Improvement of the Experimental Setup to Assess Cutaneous Bioavailability on Human Skin Models: Dynamic Protocol

Dreher

Patouillet

Fouchard

et al. 2002

Skin Pharmacol Physiol

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Human skin models, such as EpiDerm^® and Episkin^®, are not easily mounted into static or dynamic diffusion cells that are commonly used to perform bioavailability studies with human skin ex vivo. For various reasons, such as fragility, small sample size, and other morphological constraints, skin absorption studies with human skin models are often carried out on the delimited skin surface obtained by gluing a ring onto the reconstituted epidermis and manually exchanging the receptor solution. However, such an experimental setup is prone to artifacts. Discontinuous removal of the receptor fluid leads to alternating sink conditions, and an area of application smaller than the area in contact with the receptor fluid, as well as imperfect seal of the glued ring, may result in inaccurate penetration rates. Human skin models were shown to be relatively easily mounted into In-Line cells (PermeGear Inc.), vertical diffusion cells which appear to be appropriately designed for such a purpose. In-Line cells allowed accurate determination of solute penetration as well as automated sampling of receptor fluid. Excised human skin can be mounted into these cells as well, making it possible to compare penetration rates through different types of skin samples under identical conditions. Using mannitol as a reference compound, penetration profiles and epidermal distribution similar to those obtained with human skin ex vivo were obtained both with EpiDerm and Episkin. Under the present conditions, human skin models were more permeable to mannitol than excised human skin, which was only slightly permeable to mannitol. Due to these experimental innovations and to the good agreement with the absorption characteristics through human skin ex vivo, EpiDerm and Episkin seem to be promising human skin models for testing the cutaneous bioavailability of topical products in vitro.

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“…Despite the altered and unlimited growth potential, HaCaT cells, similar to normal keratinocytes, reform an orderly structured and differentiated epidermal tissue when transplanted onto nude mice (30). Hence, HaCaT cells have proved to be useful to determine, inter alia, skin toxicity or irritancy of various agents (31)(32)(33)(34)(35), to investigate the mechanisms of cutaneous allergic reactions, or of cell interactions in inflammatory, or neoplastic processes (36), as well as effects of UV-irradiation and reactive oxygen species (37)(38)(39). Next to the comparison to HaCaT cells we investigated the influence of age, gender, and skin localization on the proteome of primary human keratinocytes.…”

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confidence: 99%

Consistency of the Proteome in Primary Human Keratinocytes With Respect to Gender, Age, and Skin Localization

Sprenger

Weber

Zarai

et al. 2013

Molecular & Cellular Proteomics

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Keratinocytes account for 95% of all cells of the epidermis, the stratified squamous epithelium forming the outer layer of the skin, in which a significant number of skin diseases takes root. Immortalized keratinocyte cell lines are often used as research model systems providing standardized, reproducible, and homogenous biological material. Apart from that, primary human keratinocytes are frequently used for medical studies because the skin provides an important route for drug administration and is readily accessible for biopsies. However, comparability of these cell systems is not known. Cell lines may undergo phenotypic shifts and may differ from the in vivo situation in important aspects. Primary cells, on the other hand, may vary in biological functions depending on gender and age of the donor and localization of the biopsy specimen. Here we employed metabolic labeling in combination with quantitative mass spectrometry-based proteomics to assess A431 and HaCaT cell lines for their suitability as model systems. Compared with cell lines, comprehensive profiling of the primary human keratinocyte proteome with respect to gender, age, and skin localization identified an unexpected high proteomic consistency. The data were analyzed by an improved ontology enrichment analysis workflow designed for the study of global proteomics experiments. It enables a quick, comprehensive and unbiased overview of altered biological phenomena and links experimental data to literature. We guide through our workflow, point out its advantages compared with other methods and apply it to visualize differences of cell lines compared with primary human keratinocytes. Molecular &

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Episkin, a reconstituted human epidermis for assessing in vitro the irritancy of topically applied compounds

Cited by 70 publications

References 20 publications

Improvement of the Experimental Setup for Skin Absorption Screening Studies with Reconstructed Skin EPISKIN®

Improvement of the Experimental Setup for Skin Absorption Screening Studies with Reconstructed Skin EPISKIN®

Improvement of the Experimental Setup to Assess Cutaneous Bioavailability on Human Skin Models: Dynamic Protocol

Consistency of the Proteome in Primary Human Keratinocytes With Respect to Gender, Age, and Skin Localization

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