Thymomas, especially epithelial component were cultured, and differentiation between invasive and noninvasive thymoma was attempted.
Materials were obtained at operations and were immediately minced. A portion was digested with 500-1000 u/m
lof dispase, 0.25% trypsin or 0.05% collagenase and the remainder was placed in plastic tissue culture dishes, some of which were coated with type I collagen. Culture media were Dulbecco's modified MEM, Ham's F12 or Medium 199. These were supplemented with 10-20% fetal bovine serum, and some as the following additives were tested : 0.5 mg of ATP, 5ug/ml of dibutyryl cyclic AMP, 5 half-maximal units of IL-2, 0.05 u/m
l of Insulin, 5 ng/m
l of EGF, 10 ng/m
l of phorbol 12-myristate 13-acetate, 10
-9 M of chorela toxin, 2.5 mg/m
l of methylprednisolone, autologous peripheral blood lymphocytes.
Tissues were obtained from 9 patients with invasive and 7 with noninvasive thymoma. None of them was established and no additives promoted cell proliferation under the conditions described in this paper. Primary cultures from the invasive thymomas grew well and many epithelial cells were easily obtained. Tumor cells were dissociated easily by proteases or by physical forces. On the contrary, few or no tumor cells were obtained in the noninvasive cases. The growth of mesenchymal cells was always more dense in the noninvasive than in the invasive cases. Microscopic findings in the invasive cases were : dense cytoplasm and thick margin of nucleus, large nucleus, 1 or 2 clearly visible large nucleoli, sharply defined cell borders.
Although we cannot necessarily distinguish between invasive and noninvasive thymoma from these results alone, this method may become a useful tool in the diagnosis of thymoma.
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