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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Urological research 22 (1994), S. 67-74 
    ISSN: 1434-0879
    Keywords: Urothelium ; Primary cell culture ; Defined medium ; Epidermal growth factor ; Cytokeratins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aim of this study was to culture human urothelium and generate enough cells for subsequent reconstructive surgery. Using a modification of the Rheinwald-Green method for the routine culture of keratinocytes from patients with burns, we successfully cultured 91% of 57 biopsies from the renal pelvis, ureter, bladder and urethra of paediatric patients. The cells could be split one to three up to 9 times at 7–10 day intervals, giving a surface area of 1000 cm2 after a 2 month culture period. Primary cultures could not be initiated in defined medium MCDB153, although cells initiated using the Rheinwald-Green method could subsequently be propagated in this medium. Cytokeratin patterns in vitro were similar to those in vivo in the expression of keratins 7, 18 and 19 (characteristic of simple epithelia) and keratin 13 (characteristic of non-cornified stratified epithelia). Cultured urothelium also expressed keratin 14 (characteristic of cornified stratified epithelium) in about 25% of cells and keratin 16 (characteristic of fast-growing cells). These findings indicate that urothelial cells can be propagated in vitro for autologous grafting, and the next step is to identify substrates suitable for urothelial cell growth and differentiation and surgical manipulation.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-069X
    Keywords: Cytokeratins ; Epidermoid and trichilemmal cysts ; Milia ; Hydrocystoma ; Syringoma ; Pilomatricoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The differentiation state of skin epithelial cysts and some uncommon types of epithelial skin tumours was investigated by immunohistochemical staining, mainly using cytokeratin (CK) polypeptide-specific monoclonal antibodies. Samples of interfollicular epidermis, hair follicles and eccrine sweat glands were included as reference tissues. The CK reactivity in epidermoid cysts and milia is not restricted to CKs involved in epidermal-type differentiation, i.e. CK1, 5, 10 and 14, but in addition CK16, a hyperproliferative keratinocyte marker is suprabasally expressed. CK1 and 10 are other prominent suprabasal markers, while CK14 seems to be preferentially expressed in the basal cell layer. Of the non-epidermal CKs, only CK4 was focally or more extensively detected in about 50% of the cases. In terms of CK reactivity, keratinization of trichilemmal cysts corresponds to the keratinization of the anagen-phase hair follicle in the isthmus. The CK reactivity is again restricted to CK1, 5, 10, 14 and 16. However, the CK1 as well as CK10 reactivity is subject to serious limitations, since both CKs were only convincingly observed in foci of terminal differentiation. Eccrine hydrocystoma obligatorily expresses a complex CK set, including CK7, 8, 14, 18 and 19. This CK set perfectly corresponds to the CK composition observed in acini of eccrine sweat glands. In addition, a discontinuous CK4 and 16 reactivity was seen in about 50% of the sites, while CK1 and 10 displayed a strictly focal appearance. On the other hand, syringoma produces in its distinct structures, a CK pattern reminiscent of the one observed in eccrine sweat gland ducts and includes CK1, 5, 10, 14, 16 and 19. Finally, the CK expression pattern of pilomatricoma includes CK1, 8, 10, 14 and 19, and is reminiscent of the CK staining of hair bulb matrix cells differentiating in the keratogenous zone in the direction of hair cortex. The reactivity of CK1 and 10 was mainly restricted to foci of squamoid differentiation and also to transitional cells bordering on shadow cells, as far as it concerns CK10. Occasionally, CK7 and 16 were observed in individual cells or small cell groups. In our view these CK reactivity patterns are useful to judge the differentiation state reached in pathological conditions, but so far do not allow us unequivocally to determine the site of origin of these lesions.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-069X
    Keywords: Seborrhoeic keratoses ; Cytokeratins ; Epidermal transglutaminase ; Involucrin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The mode of differentiation of seborrhoeic keratoses was investigated by immunohistochemical staining using cytokeratin (CK) polypeptide-specific monoclonal antibodies and an antibody specific for the particulate form of epidermal transglutaminase (ETgase), and by applying an anti-human involucrin serum. The role played by (E)Tgase was further evaluated using an activity assay based on the covalent attachment of monodansylcadaverine. Samples of uninvolved epidermis served as reference tissue. CK reactivities suggested that seborrhoeic keratoses is a hyperproliferative disease with an epidermal CK composition. CK5 and CK14 were prominent markers of basal and basaloid keratinocytes, whereas a decrease in staining occurred in advanced maturation stages and areas of terminal keratinization. In contrast, CK1 and CK10 were prominent markers of suprabasaloid differentiation stages and produced complementary stainings to those of CK5 and 14. Generally, CK10 staining was more impressive than CK1 staining and seemed to start before CK1 staining. In contrast to CK10 staining, cornified areas lost CK1 reactivity. These staining patterns were similar to those observed in uninvolved reference tissues. The epidermal CK subset was further supplemented with the ‘hyperproliferative’ CK6 and 16 which occur sequentially. Positive staining for CK6 was noted from basal and proximal basaloid cells onwards, whereas distal basaloid cells additionally showed CK16 staining. The presence of other non-epidermal CK polypeptides could not be shown. The competence for other differentiation markers belonging to the group of (E)Tgase and cornifying cell membranes also evolved with a typical epidermal pattern. (E)Tgase activity was restricted to advanced and terminal stages of keratinization and was dual in nature, i.e. a diffuse cytoplasmic staining occurred together with a prominent staining of cornifying cell membranes. Similarly, involucrin first detected in the cytosol of distal basaloid cells, was soon translocated to the cornifying cell membrane, reflecting its function as an ETgase substrate and precursor of the marginal band. Finally, the immunolocalization of the particulate form of ETgase was strikingly similar to the location of the first two markers. Taken together, the results allow us to conclude that seborrhoeic keratoses exhibits a hyperproliferative variant of the epidermal keratinization process. Maturation of basal keratinocytes is greatly retarded leading to an accumulation of basaloid cells which retain the molecular markers of basal cells in proximal areas, but progressively gain the molecular markers of advancing maturation in distal areas.
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