Histomorphological and biochemical differentiation capacity in organotypic co-cultures of primary gingival cells
| Autor: | Gerda Komposch, N. E. Fusenig, Annette Kohl, Pascal Tomakidi |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Keratinocytes
Pathology medicine.medical_specialty Keratin 14 Epithelial Attachment Gingiva Gene Expression Vimentin Biology Basement Membrane Epithelium Mesoderm Laminin Keratin medicine Humans Fluorescent Antibody Technique Indirect Cells Cultured In Situ Hybridization Tissue homeostasis chemistry.chemical_classification Basement membrane Cell Differentiation Epithelial Cells Fibroblasts Molecular biology Coculture Techniques medicine.anatomical_structure chemistry Organ Specificity Cell culture biology.protein Keratins Periodontics Collagen |
| Zdroj: | Journal of Periodontal Research. 32:388-400 |
| ISSN: | 1600-0765 0022-3484 |
| DOI: | 10.1111/j.1600-0765.1997.tb00549.x |
| Popis: | To establish a three-dimensional in vitro test system mimicking the physiological situation of the oral cavity, organotypic co-cultures consisting of primary gingival cells on a collagen matrix with fibroblasts were generated. The histomorphological development after 7 and 14 d revealed close similarity with the non-keratinized gingiva epithelium. Furthermore, as epithelial specific markers synthesis and localization of keratins as well as the deposition of basement membrane components were assessed on frozen sections by immunofluorescence and keratin expression by in situ hybridization. Primary keratinocytes in conventional culture strained positive for keratin K14 and the mucosal differentiation-specific keratins K4 and K13, while primary fibroblasts, isolated from the same tissue source, and also some keratinocytes, were positive for vimentin. In organotypic co-cultures the keratinocytes formed a multilayered epithelium within 14 d containing basal cells and flattened cells in the uppermost layers. Comparable to native non-keratinized gingiva keratin 14 gene expression was clearly detectable in the basal cell compartment but showed extending immunolocalization. In addition, particularly at the early stage (7 d), basally located keratinocytes were also vimentin positive. According to morphological differentiation K4 and K13 were detectable in suprabasal position a the RNA and protein level. The major basement membrane constituents collagen type IV and laminin increased with time revealing first an interrupted and later a fully extended staining underneath the basal cells. Maintenance of basal cell function was further demonstrated by cell proliferation (BrdU incorporation) which was initially high (7 d) but declined towards the later stages (14-21 d). The results demonstrate i) that this co-culture system leads to a stratified surface epithelium with morphological and biochemical characteristics of the non-keratinized gingiva epithelium and ii) that a state of physiological tissue balance was reached, thus rendering a suitable model for tissue compatibility studies. |
| Databáze: | OpenAIRE |
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