Anti-pan Cytokeratin monoclonal antibody, clone D-12

Background: Mammal skin plays a pivotal role in several life preserving processes and extensive damage may therefore be life threatening. Physiological skin regeneration is achieved through ongoing somatic stem cell differentiation within the epidermis and the hair follicle. However, in severe pathological cases, such as burn wounds, chronic wounds, and ulcers, the endogenous repair mechanisms might be insufficient. For this reason, exogenous purification and multiplication of epithelial-like stem/progenitor cells (EpSCs) might be useful in the treatment of these skin diseases. However, only few reports are available on the isolation, purification and characterization of EpSCs using suspension cultures. Methods: In the present study, skin was harvested from 6 mares and EpSCs were isolated and purified. In addition to their characterization based on phenotypic and functional properties, sphere formation was assessed upon isolation, i.e. at passage 0 (P-0), and at early (P-4) and late (P-10) passages using different culture conditions. Results: On average 0.53 +/- 0.28% of these primary skin-derived cells showed the capacity to form spheres and hence possessed stem cell properties. Moreover, significantly more spheres were observed in EpSC medium versus differentiation medium, corroborating the EpSCs' privileged ability to survive in suspension. Furthermore, the number of cells per sphere significantly increased over time as well as with subsequent passaging. Upon immunophenotyping, the presumed EpSCs were found to co-express cytokeratin (CK) 14, Casein kinase 2 beta and Major Histocompatibility Complex (MHC) I and expressed no pan CK and wide CK. Only a few cells expressed MHC II. Their differentiation towards keratinocytes (at P-4 and P-10) was confirmed based on co-expression of CK 14, Casein kinase 2 beta, pan CK and wide CK. In one of six isolates, a non-EpSC cell type was noticed in adherent culture. Although morphological features and immunohistochemistry (HC) confirmed a keratinocyte phenotype, this culture could be purified by seeding the cells in suspension at ultralow clonal densities (1 and 10 cells/cm(2)), yet with a significantly lower sphere forming efficiency in comparison to pure EpSCs (P = 0.0012). Conclusion: The present study demonstrated sphere formation as a valuable tool to purify EpSCs upon their isolation and assessed its effectiveness at different clonal seeding densities for eliminating a cellular contamination. Copyright (C) 2014 S. Karger AG, Basel

A significant fraction of nasopharyngeal and sinonasal tumors are associated with Epstein-Barr virus (EBV) or human papillomavirus (HPV). Nasopharyngeal carcinoma (NPC) and extranodal NK/T-cell lymphoma harbor EBV in practically all cases, although a small proportion of cases of the former harbor HPV. Sinonasal inverted papillomas harbor HPV in about 25% of cases. Sinonasal squamous cell carcinomas harbor transcriptionally active HPV in about 20% of cases, and limited data suggest that this subset has a better prognosis than the HPV-negative subset. This review addresses the diagnostic issues of the EBV-associated tumors. Difficulties in diagnosis of NPC may be encountered when there are prominent crush artifacts, many admixed lymphoid cells masking the neoplastic cells, or numerous interspersed granulomas, whereas benign cellular components (epithelial crypts and germinal centers) and reactive lymphoid hyperplasia can potentially be mistaken for NPC. Immunostaining for pan-cytokeratin and/or in situ hybridization for EBER can help in confirming or refuting a diagnosis of NPC. The main diagnostic problem of extranodal NK/T-cell lymphoma is recognition of the neoplastic nature of those examples predominated by small cells or showing a mixture of cells. The identification of a destructive infiltrate (dense expansile infiltrate; angiocentric growth) and definite cytologic atypia (clear cells; many medium-sized cells) would favor a diagnosis of lymphoma, which can be supported by immunohistochemistry (most commonly CD3+, CD5-, CD56+) and in situ hybridization for EBER. In conclusion, among nasopharyngeal and sinonasal neoplasms, demonstration of EBV may aid in diagnosis, particularly NPC and extranodal NK/T-cell lymphoma. Demonstration of HPV does not have a role yet in diagnosis, although this may change in the future.