Epidermal Organization and Differentiation of HaCaT Keratinocytes in Organotypic Coculture with Human Dermal Fibroblasts

来自掌桥科研

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184

作者：

VM Schoop，N Mirancea，NE Fusenig

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摘要：

The immortal keratinocyte line HaCaT is frequently used as a paradigm for skin keratinocytes in vitro because of its highly preserved differentiation capacity. HaCaT cells form a nearly regular epidermal architecture when transplanted onto subcutaneous tissue of athymic . In order to analyze further their differentiation capacity in vitro, HaCaT cells were studied in organotypic cocultures on top of gels containing dermal fibroblasts. Within 1 wk HaCaT cells formed a still dysplastic epithelium, the thickness of which correlated with the number of fibroblasts in the gel. With further culture time of up to 3 wk a remarkably well structured and differentiated squamous epithelium developed. After 1 wk, and 16, , and I were expressed in suprabasal layers, whereas , keratin 2e, and appeared after 2-3 wk. Within this time, a nearly complete had formed including and anchoring fibrils. became restricted to the basal layer after 2 and 3 wk. Using the TdT-mediated nick end labeling assay, fragmentation of DNA was detectable in nuclei of the parakeratotic stratum corneum. Ultrastructurally, many features of keratinization accumulated after 2 and 3 wk, though an orthokeratotic keratinization was not achieved, in contrast to HaCaT transplants. This differentiation deficiency - as compared with normal keratinocytes -- might be due to a lack of paracrine factors important for or to a reduced sensitivity of these cells. Nevertheless, this high degree of differentiation under organotypic conditions qualifies this cell line as an appropriate model for elucidation of the molecular mechanisms regulating keratinocyte growth and differentiation and for use in pharmacotoxicology.

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关键词：

Humans Basement Membrane Epidermis Fibroblasts Cell Line Biological Markers Coculture Techniques Cell Communication Cell Division Cell Differentiation