Immunobead and Density Gradient Purification of Paget Cells:In vitroStudies of Proliferation
| Autor: | Takashi Hashimoto, Minoru Miyasato, Osamu Mori, Tadashi Karashima |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Male
musculoskeletal diseases Pathology medicine.medical_specialty Density gradient medicine.drug_class Iohexol Paget's Disease Mammary Contrast Media Breast Neoplasms Cell Count Dermatology In Vitro Techniques Inhibitory postsynaptic potential Monoclonal antibody Carcinoembryonic antigen Tumor Cells Cultured otorhinolaryngologic diseases medicine Humans Aged Aged 80 and over Differential centrifugation Epidermis (botany) biology Mucin-1 General Medicine Molecular biology In vitro Carcinoembryonic Antigen Paget Disease Extramammary Microscopy Fluorescence Cell culture Genital Neoplasms Male biology.protein Female |
| Zdroj: | The Journal of Dermatology. 25:497-502 |
| ISSN: | 0385-2407 |
| DOI: | 10.1111/j.1346-8138.1998.tb02443.x |
| Popis: | Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramamary Paget's disease investigated whether Paget cells proliferate as other malignant cells do. Although epithelial monolayers from the involved skin were maintained for approximately 45 days, no permanent cell lines were established. The proportion of carcinoembryonic antigen (CEA)-positive cells did not increase in the long-term cultures. Herein, we report studies of whether there is a real reduction of Paget cell numbers or if this is merely a decrease in the expression of CEA by the cells. Furthermore, we investigated whether Paget cells survive longer when cultured free from any potential inhibitory keratinocytes or other epidermal cells. Skin samples were obtained from one patient with mammary Paget's disease and three with extramammary Paget's disease; epidermal cells were cultured in vitro. An enrichment of Paget cells was carried out from the cultured epidermal cells by combining an antiepithelial membrane antigen monoclonal antibody, binding to immunobeads, and density gradient centrifugation in Nycodenz. The separated cells were re-cultured in Keratinocyte-SFM serum-free media. The proportion of CEA-positive cells did not increase in the culture, and the purified cells did not show any increase in survival times compared to the non-purified cultured cells. These results suggest that the decrease of CEA-positive cells noted during culture results from a decline in expression of CEA in the Paget cells. Paget cells in the involved epidermis do not proliferate significantly and thus differ from many other malignant cells. |
| Databáze: | OpenAIRE |
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