Comparative phenotypic characterization of keratinocytes originating from hair follicles |
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Authors: | Jiří Klíma Karel Smetana Jr Jan Motlík Zuzana Plzáková Fu-Tong Liu Jiří Štork Herbert Kaltner Martin Chovanec Barbora Dvořánková Sabine André Hans-Joachim Gabius |
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Institution: | (1) Institute of Animal Physiology and Genetics, Academy of Sciences, Libechov, Czech Republic;(2) 2nd Faculty of Medicine, Center of Cell Therapy and Tissue Repair, Charles University, Prague, Czech Republic;(3) 1st Faculty of Medicine, Institute of Anatomy, Charles University, Prague, Czech Republic;(4) School of Medicine, Department of Dermatology, University of California, Davis,Sacramento, CA USA, 95817;(5) 1st Faculty of Medicine, Department of Dermatovenerology, Charles University, Prague, Czech Republic;(6) Faculty of Veterinary Medicine, Institute of Physiological Chemistry, Ludwig-Maximilians-University, Munich, Germany;(7) 1st Faculty of Medicine, Department of Otorhinlaryngology, Head and Neck Surgery, Charles University, Prague, Czech Republic;(8) 3rd Faculty of Medicine, Department of Burn Surgery, Charles University, Prague, Czech Republic |
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Abstract: | 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 Np63 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. |
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