Does anybody have a protocol and successfully isolated, grown and expanded cancer cells from leukapheresis of patients with solid tumors such as breast, colon, pancreatic, lung cancer ?
Lombardi Cancer Center Georgetown University Washington DC
Dear Anton,
this is a difficult task. The main problem is not the isolation of the cancer cells, but that these cancer cells in most cases will not grow in culture (for unknown reasons). You may try it this way:
The situation is, e.g., a patient with breast carcinoma and very very few cancer cells in her blood and you get about 50ml peripheral blood. With Ficoll you can get rid of erythrocytes and granulocytes. Wash the buffy coat once in whole medium with heparin. Now I would avoid any further purification or concentration of the tumor cells. Cell sorting is pure stress for cells. Seed all cells in one flasc, wait until the tumor cells - and probably other cells too - have attached. Wash away floating lymphocytes. And don't be afraid if cells like macrophages, dendritic cells, or other normal cells are attached too. These cells normally dissappear after 2-3 weeks. Good luck.
Cave: It happens very often that a lymphoblastoid cell line develops spontaneously. These cells are growing agressively and they form small clumps that look somehow "hairy". They express B cell markers and have a normal karyotype. And you cannot get rid of these cells.
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