PubMed Health⌕ Search

Biomedical subjects

Jirí Stork

Publications and source records attributed to Jirí Stork.

4 recordsLinked to original sources

Immunocyto- and histochemical profiling of nucleostemin expression: marker of epidermal stem cells?

BACKGROUND AND OBJECTIVE: Because the nucleolar protein nucleostemin is present in bone marrow and neuronal stem cells and malignancies originating thereof we monitored its expression in frozen sections from normal human epidermis, basal cell carcinomas, cultured keratinocytes and cells of the squamous carcinoma line FaDu. In addition, probing the value of this protein as a marker of epidermal stem cells was an aim of this study. MATERIALS AND METHODS: To further characterize cell features we added analysis of expression of keratins 10 or 19 as markers of terminal differentiation and Ki67 as marker of proliferating cells as well as three adhesion/growth-regulatory galectins. RESULTS: Immunohistochemical monitoring revealed expression of nucleostemin in cells of both Ki67-positive and -negative nuclei regardless of the K10-expression status. Cultured keratinocytes were positive, when they were prepared from hair follicles and cultured in the presence of feeder cells. A small population of these nucleostemin-positive cells also expressed galectin-1 but not galectins-3 and -9 in their nucleoli. Part of these cells also expressed keratin 19. FaDu cells were strongly positive, illustrating expression in malignant cells which require no feeder layer. Of note, the number of galectin-1-positive nucleoli was reduced in the course of culture. CONCLUSION: Nucleostemin positivity cannot be considered as marker for stem cells in skin sections. In cultured cells, nucleostemin is expressed in a distinct population of the epidermal cells from hair follicle kept in the presence of a feeder layer, intimating an association of nucleostemin expression with this type of epithelio-mesenchymal interaction which is not essential during propagation of malignant cells.

Biomarkers↗

Comparative phenotypic characterization of keratinocytes originating from hair follicles.

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.

Animals↗

Transient expression of keratin 19 is induced in originally negative interfollicular epidermal cells by adhesion of suspended cells.

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.

Basal Cell Carcinoma↗

Granular cell basal cell carcinoma.

A 69-year-old woman presented with a 2-year history of a slowly growing tumour on the nose. The clinical diagnosis was basal cell carcinoma (BCC) and a complete excision was performed. Histologically, the tumour had the general features of a BCC but with sheets and nests of cells with granular cytoplasmic changes in the centre of the lesion. A pathological diagnosis of granular cell BCC was made. On immunohistochemical examination, the tumour cells were Ber-EP4, cytokeratin AE1/AE3 and cytokeratin CAM 5.2 positive but S100 protein negative. Only the granular cells were CD68 antigen (monoclonal antibody KP1) positive.

Adult↗