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  • Keratin 6  (1)
  • Key words Reconstructed epidermis  (1)
  • 1
    ISSN: 1432-069X
    Keywords: Key words Keratinocyte ; Keratin 6 ; EGF ; Triglyceride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In this study we compared human keratinocyte cultures grown at the air-liquid interface on de-epidermized dermis at 33° C or at 37° C in two different culture media: medium I – a fully defined serum- and EGF-free medium; and medium II – a serum- and EGF-containing medium. Cultures grown in medium II were initially hyperproliferative followed rapidly by senescence, and had a high triglyceride content. The hyperproliferation was ascribed to the presence of EGF in the medium. In contrast, cultures grown in medium I at 33° C showed a greatly improved balance between cell proliferation and differentiation. They had a prolonged lifespan of at least 32 days without a significant decrease in the number of living cell layers, a rate of proliferation similar to that of native epidermis and a low triglyceride content. Culturing at 37° C increased the rate of differentiation without affecting the rate of proliferation. Furthermore, both at 33° C and at 37° C, keratin 6 was expressed only in the first suprabasal layer but was expressed in all suprabasal layers in cultures grown in medium II. High keratin 6 expression was not directly linked to hyperproliferation but to deregulated terminal differentiation. Involucrin, transglutaminase and SPRR1 were abnormally expressed irrespective of the culture conditions used, whereas SKALP expression was decreased in cultures grown in medium I. The epidermal lipid profile was better in cultures grown in medium I; the relative amounts of ceramides, free fatty acids and cholesterol being comparable to native epidermis. Small-angle X-ray diffraction showed a slightly improved structural organization of stratum corneum lipids as demonstrated by the appearance of second- and third-order peaks of the 12-nm long phase and a marked reduction in the polycrystalline cholesterol peak.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Key words Reconstructed epidermis ; Differentiation ; Proliferation ; Migration ; Loricrin ; Involucrin ; SPRR ; EGF ; TGFα ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Epidermis reconstructed on de-epidermized dermis was used to investigate the effects of growth factors and culture temperature on epidermal morphogenesis and the expression of cornified envelope precursors. Cultures grown at 33°C or 37°C in the absence or presence of transforming growth factor alpha (TGFα), keratinocyte growth factor (KGF), basic fibroblast growth factor (bFGF), or insulin-like growth factor (IGF) show a similar morphology to that of native epidermis. Loricrin and SPRR2 are expressed in the stratum granulosum and SPRR3 is absent. Cultures grown in epidermal growth factor (EGF)-supplemented medium at 37°C have a normal morphology, whereas cultures grown at 33°C have a disorganized basal layer, no stratum granulosum, and nuclei are present in the stratum corneum. Loricrin is absent, and SPRR2 and SPRR3 expression extend into the spinous layers. Irrespective of the culture condition used, involucrin is aberrantly expressed in all suprabasal layers. EGF stimulated keratinocyte proliferation and migration to a greater degree than TGFα. Epidermis reconstructed on fibroblast-populated collagen gels at 33°C led to the same disturbances in keratinocyte differentiation as seen in cultures grown on de-epidermized dermis at 33°C in the presence of EGF, whereas parallel cultures grown at 37°C have a similar morphology to that of native epidermis.
    Type of Medium: Electronic Resource
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