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001-es BibID:BIBFORM039247
Első szerző:Havlickova, Blanka
Cím:Towards optimization of an organotypic assay system that imitates human hair follicle-like epithelial-mesenchymal interactions / Havlickova, B., Biro, T., Mescalchin, A., Arenberger, P., Paus, R.
Dátum:2004
ISSN:0007-0963
Megjegyzések:Human hair growth can currently be studied in vitro by the use of organ-cultured scalp hair follicles (HFs). However, simplified organotypic systems are needed for dissecting the underlying epithelial-mesenchymal interactions and as screening tools for candidate hair growth-modulatory agents. OBJECTIVES: To optimize the design and culture conditions of previously published organotypic systems that imitate epithelial-mesenchymal interactions in the human HF as closely as possible. MATERIALS AND METHODS: Continuous submerged organotypic 'sandwich' cultures were established. These consist of a pseudodermis (collagen I mixed with and contracted by human interfollicular dermal fibroblasts) on which one of two upper layers is placed: either a mixture of Matrigel basement membrane matrix (BD Biosciences, Bedford, MA, U.S.A.) and follicular dermal papilla fibroblasts (DPC), with outer root sheath keratinocytes (ORSK) layered on the top ('layered' system), or a mixture of Matrigel, DPC and ORSK ('mixed' system). Morphological and functional characteristics of these 'folliculoid sandwiches' were then assessed by routine histology, histomorphometry and immunohistochemistry. RESULTS: In both 'layered' and 'mixed' systems, the ORSK formed spheroid epithelial cell aggregates, which retained their characteristic keratin expression pattern (i.e. cytokeratin 6). In the 'mixed' sandwich model the size of the epithelial cell aggregates was smaller, but the numbers of ORSK were significantly higher than in the 'layered' model at day 14 in the culture. ORSK proliferated better in the 'mixed' than in the 'layered' sandwich system, regardless of the calcium or serum content of the media, whereas apoptosis of ORSK was lowest in the 'mixed' system in serum-free, low calcium medium. The kinetics of proliferation and apoptosis of DPC, which retained their characteristic expression of versican, were similar in both systems. However, proliferation and apoptosis of DPC were higher in the presence of serum and/or under high calcium conditions. CONCLUSIONS: Our results underscore the importance of structural design and medium composition for epithelial-mesenchymal interactions as they occur in the human HF. Specifically, we report a new organotypic submerged 'folliculoid sandwich' system with serum-free, low calcium medium and a mixture of interacting human DPC and ORSK, which offers several advantages over previously available assays. This system allows the standardized assessment of the effects of a test agent on the proliferation, apoptosis and key marker expression of human ORSK and DPC under substantially simplified in vitro conditions which approximate the in vivo situation.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:British Journal Of Dermatology. - 151 : 4 (2004), p. 753-765. -
További szerzők:Bíró Tamás (1968-) (élettanász) Mescalchin, Alessandra Arenberger, P. Paus, Ralf
Internet cím:Szerző által megadott URL
DOI
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2.

001-es BibID:BIBFORM009198
Első szerző:Havlickova, Blanka
Cím:A human folliculoid microsphere assay for exploring epithelial- mesenchymal interactions in the human hair follicle / Havlickova, B., Biro, T., Mescalchin, A., Tschirschmann, M., Mollenkopf, H., Bettermann, A., Pertile, P., Lauster, R., Bodo, E., Paus, R.
Dátum:2009
ISSN:1523-1747 (Electronic)
Megjegyzések:The search for more effective drugs for the management of common hair growth disorders remains a top priority, both for clinical dermatology and industry. In this pilot study, we report a pragmatic organotypic assay for basic and applied hair research. The patented technique produces microdroplets, which generate human folliculoid microspheres (HFMs), consisting of human dermal papilla fibroblasts and outer root sheath keratinocytes within an extracellular matrix that simulates elements of the hair follicle mesenchyme. Studying a number of different markers (for example, proliferation, apoptosis, cytokeratin-6, versican), we show that these HFMs, cultured under well-defined conditions, retain several essential epithelial-mesenchymal interactions characteristic for human scalp hair follicle. Selected, recognized hair growth-modulatory agents modulate these parameters in a manner that suggests that HFMs allow the standardized preclinical assessment of test agents on relevant human hair growth markers under substantially simplified in vitro conditions that approximate the in vivo situation. Furthermore, we show by immunohistochemistry, reverse transcriptase-PCR, and DNA microarray techniques that HFMs also offer a useful discovery tool for the identification of target genes and their products for candidate hair drugs. HFM thus represent an instructive modern experimental and screening tool for basic and applied hair research in the human system.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Apoptosis
Cell Communication
Cell Proliferation
Cells, Cultured
Cyclosporine
Cytokines
Epithelial Cells
Gene Expression Profiling
Hair Follicle
Hepatocyte Growth Factor
Humans
Mesoderm
Microspheres
Megjelenés:The Journal of Investigative Dermatology. - 129 : 4 (2009), p. 972-983. -
További szerzők:Bíró Tamás (1968-) (élettanász) Mescalchin, Alessandra Tschirschmann, Miriam Mollenkopf, Hans Bettermann, Albrecht Pertile, Paolo Lauster, Roland Bodó Enikő Paus, Ralf
Internet cím:DOI
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