Using cancer cells (adenoma) from ascites in three gastric cancer patients and tumor cells from ascites in three cases with coelothelioma the author conducted tissue culture of these cells in suspension by roller-tubes and further studied morphological changes of these tumor cells during the culture under a phase-contrast-microscope, and obtained the following results.
1) As for the method of tissue culture, it is most suitable to use the medium composed of 70 per cent Hank's fluid, 15 per cent chick embryo extract, 15 per cent normal human serum or serum of various cancer patients, with the addition of 2 mg/cc ribonucleic acid and vitamin B12 2 γ/cc. At the same time ground chick embryo tissue is cultured with this cellsuspension culture. Moreover, the medium is exchanged with fresh one every three days and the ground chick embryo tissue every six days. 2) By this culture method it has been possible to keep the cancer cells of ascites in gastric cancer alive for 21 days while tumor cells of coelothelioma for 18 days. 3) In the morphological observations: (1) In the case of cancer cells of gastric cancer ascites, cell division increases around third to fourth day of culture and numerous new small cells with clear cell body appear. On the other hand, about this time a portion of the cells become degenerated and are destroyed and within 4 or 5 days as a cyclic period the processes of cell degeneration and destruction and appearance of regenerated small cells are repeated. (2) Likewise in the tissue culture of coelothelioma cells, cell division occurs on around the third day of culture, and new small cells with clear cell-body make their appearance, revealing distinctly the transformation from serous cells. By the ninth day the cells gradually become disintegrated and are destroyed, and with lapse in culture time degenerated cells increase in number. As can be seen from above, it has been possible to obtain quite interesting findings in the day-to-day observations on morphological changes occurring in tumor cells under culture by the above method.