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Immunobead and Density Gradient Purification of Paget Cells: In vitro Studies of Proliferation
Author(s) -
Mori Osamu,
Miyasato Minoru,
Karashima Tadashi,
Hashimoto Takashi
Publication year - 1998
Publication title -
the journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.9
H-Index - 65
eISSN - 1346-8138
pISSN - 0385-2407
DOI - 10.1111/j.1346-8138.1998.tb02443.x
Subject(s) - epidermis (zoology) , carcinoembryonic antigen , in vitro , cell culture , antigen , monoclonal antibody , keratinocyte , cell , biology , pathology , antibody , microbiology and biotechnology , cancer research , immunology , medicine , cancer , anatomy , biochemistry , genetics
Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramammary 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 anti‐epithelial 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 cultures, 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.