Keratin 19 and nuclear reactivity to an endogenous lectin, galectin-1, represent a potential marker of epidermal stem cells. We detected expression of keratin 19 and nuclear binding sites for galectin-1 in adult cells migrating from the hair follicle, where cells expressing keratin 19 are located in the bulge region. The results were compared with the expression of both markers in cells adhering from suspension prepared from the interfollicular epidermis without keratin-19-positive cells and with nuclear binding sites for galectin-1. The results were compared with data from basal cell carcinomas. All cells were analyzed concerning size, as it is known that cell diameter influences the clonogenic potential of keratinocytes. The major result of this study is the observation of transient expression of keratin 19 and nuclear galectin-1 binding sites in originally negative interfollicular epidermal cells induced by adhesion. These cells were very small in size, similar to basal cells of the interfollicular epidermis or the bulge region of the hair follicle. The influence of the suspension regimen on beta1-integrin expression, cell diameter and growth was also monitored. A population of cells highly positive for beta1 integrin of the same diameter as keratin-19-positive cells insensitive to induction of terminal differentiation by lack of anchorage was characterized. Cells of the same size were also observed in the keratin-19-positive cells of basal cell carcinomas. In conclusion, the expression of poor levels of differentiation induced by cell adhesion is transient. Also, keratin 19 expression should not be exclusively regarded as a marker of stem cell activity.

• MeSH
• antigeny CD29 analýza   MeSH
• bazocelulární karcinom metabolismus   patologie   MeSH
• buněčná adheze MeSH
• buněčné kultury MeSH
• časové faktory MeSH
• epidermální buňky MeSH
• epidermis chemie   MeSH
• galektin 1 analýza   MeSH
• galektin 3 analýza   MeSH
• keratin-10 MeSH
• keratin-20 MeSH
• keratinocyty chemie   cytologie   MeSH
• keratiny analýza   MeSH
• kultivované buňky MeSH
• lidé MeSH
• nádory kůže metabolismus   patologie   MeSH
• pohyb buněk MeSH
• proteiny intermediálních filament analýza   MeSH
• vlasový folikul chemie   cytologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• antigeny CD29 MeSH
• galektin 1 MeSH
• galektin 3 MeSH
• keratin-10 MeSH
• keratin-20 MeSH
• keratiny MeSH
• KRT10 protein, human MeSH Prohlížeč
• KRT20 protein, human MeSH Prohlížeč
• proteiny intermediálních filament MeSH

The principal pool of epidermal stem cells is located in the bulge region of the hair follicle root sheath. In this research project, we have used a refined procedure to isolate porcine hair follicles including their root sheath and for comparison purposes also human cell material. These cells migrating from the hair follicles were then cytochemically characterized. A panel of antibodies and two labeled plant lectins were tested on cell material obtained under a range of assorted experimental conditions. Due to their role in growth regulation we also studied two endogenous lectins, specifically monitoring their expression and the presence of accessible ligands. These in vitro results were compared with findings on porcine and human hair follicles and human basal cell carcinomas in situ. The keratinocytes originating from hair follicles in the presence of feeder cells are rather undifferentiated and express galectin-1/galectin-1-binding sites but not galectin-3 in their nuclei associated with DeltaNp63alpha positivity. Nuclear reactivity for galectin-1 was rarely observed in the bulge of the outer root sheath of the human hair follicle and of basal cell carcinomas and absent in porcine tissue samples. Exclusion of feeder cells from our cultivation system of porcine hair follicles led to the formation of spheroid bodies from these keratinocytes. Ki67 as a marker of proliferation was not present in the nuclei of cells forming these spheroids. One part of these bodies is positive for markers of post-mitotic differentiated cells, while the other spheroids are composed of poorly differentiated cells, which are able to adhere to feeder cells and form growing colonies. In summary, the detection of galectin-1 and also nuclear binding sites for this endogenous effector points to intracellular functionality of this lectin. It can be considered a potential marker of a distinct cell population, probably at the beginning of a differentiation cascade of keratinocytes.

• MeSH
• buněčná diferenciace MeSH
• fenotyp MeSH
• galektiny analýza   MeSH
• imunodominantní epitopy analýza   MeSH
• keratinocyty chemie   MeSH
• lidé MeSH
• prasata MeSH
• vlasový folikul chemie   cytologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• galektiny MeSH
• imunodominantní epitopy MeSH

A basal layer of squamous epithelia such as epidermis contains stem cells, transit amplifying cells as well as postmitotic differentiating cells. A detailed knowledge of the transition among these cell types in the course of epidermal renewal is important. It would help in better understanding of many pathological processes, including cancer, and in employment of epidermal cells for therapeutic purposes. In this study we analyzed the possible role of Dolichos biflorus agglutinin (DBA)-reactive alpha-N-acetylgalactosamine glycosylation in behavior of the human epidermal basal cells under in vivo and in vitro conditions. The data received from porcine epidermis were also included. Part of basal cells was positive for DBA-binding sites and these cells exhibited a lower presence of beta1 integrin in their basal surface connected to the basement membrane. The perinuclear Golgi-like accumulation of beta1 integrin was observed in some cultured keratinocytes. The co-localization of integrin with DBA-binding sites and 58 kDa protein suggests that alpha-N-acetylgalactosamine glycosylation could be related to beta1 integrin retention in the endoplasmatic reticulum Golgi intermediate compartment (ERGIC) at the beginning of the secretory pathway. The lack of anchorage in culture elevated the number of DBA-binding site positive cells without significant influence on cell growth when cells isolated directly from epidermis were employed in study. Some role of DBA-reactive glycoligand expressions in a suprabasal movement of differentiated basal cells can be hypothesized.

• MeSH
• acetylgalaktosamin metabolismus   MeSH
• antigeny CD29 analýza   MeSH
• buněčná adheze MeSH
• buněčná diferenciace MeSH
• dospělí MeSH
• druhová specificita MeSH
• epidermální buňky MeSH
• glykosylace MeSH
• keratinocyty chemie   cytologie   MeSH
• lidé MeSH
• plod MeSH
• pohlavní dimorfismus MeSH
• prasata MeSH
• rostlinné lektiny metabolismus   MeSH
• vazebná místa MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylgalaktosamin MeSH
• antigeny CD29 MeSH
• dolichos biflorus agglutinin MeSH Prohlížeč
• rostlinné lektiny MeSH

RHPS, composed of confluent allogeneic keratinocytes cultured on cell-free pig dermis, stimulates wound healing when applied with the keratinocyte layer facing the wound. So far it has not been clarified whether the confluent keratinocytes implanted 'upside-down' can 'take' or only stimulate healing by producing growth factors. Confluent male keratinocytes were grafted onto donor sites of three female patients. Biopsies were taken on days 4, 6 and 9 after grafting. The fate of donor cells was followed in paraffin sections by FISH for the Y chromosome and by persisting expression of vimentin taken as a marker of cultured keratinocytes. Histological evaluation was complemented by detection of keratin 10 and involucrin. All three donor sites healed within one week. On day 4 the early neoepidermis was multilayered but disordered after transplantation. A large proportion of cells were apparently of donor origin as indicated by the presence of Y chromosomes, irregular morphology, expression of vimentin in the bottom and upper layers of the neoepidermis, and by irregular expression of involucrin and keratin 10 only in the central layer of the neoepidermis. From day 6 onwards, the new epidermis acquired an ordered stratification. Involucrin and keratin 10 renewed normal distribution in suprabasal layers. Concomitantly, vimentin expression was decreasing. The Y chromosome was still found on day 6 but not on day 9. We concluded that confluent allogeneic keratinocytes temporarily 'take' to the wound and contribute to rapid wound closure, being replaced by the patient's epidermal cells after about one week.

• MeSH
• biologické markery MeSH
• chromozom Y * MeSH
• dospělí MeSH
• hojení ran MeSH
• homologní transplantace patologie   MeSH
• hybridizace in situ fluorescenční MeSH
• keratin-10 MeSH
• keratinocyty chemie   transplantace   MeSH
• keratiny analýza   MeSH
• kokultivační techniky MeSH
• kultivované buňky chemie   transplantace   MeSH
• lidé MeSH
• popálení chirurgie   MeSH
• prasata MeSH
• přežívání štěpu MeSH
• proteinové prekurzory analýza   MeSH
• škára MeSH
• tkáňové inženýrství MeSH
• transplantáty MeSH
• viabilita buněk MeSH
• vimentin analýza   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• involucrin MeSH Prohlížeč
• keratin-10 MeSH
• keratiny MeSH
• KRT10 protein, human MeSH Prohlížeč
• proteinové prekurzory MeSH
• vimentin MeSH