FRUKTO細胞の増殖と同調培養法

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The synchronizing technique which imposes a minimal stress on the system was first reported by Terasima and Tolmach. This technique has been extended to another cell line, FRUKTO cells, which was derived from mouse sarcoma (TAKIZAWA). The FRUKTO cells #22 have been grown in the medium of 5% calf serum-N16SF. The average doubling time of cells in this medium is 26.1 hr, and duration of G_1, S, G_2 and M period are estimated to be 7.4 hr,- 12.5 hr, 4.5 hr and 0.7 hr. The culture starts in the cell number of 1 x 10^5 introduced in a 180ml-squre bottle. After 40 hr of incubation the culture is agitated by the hitting apparatus which was newly devised by the author for the purpose of harvesting. One minute of powerful agitation was found necessary to collect mitotic cells apart from the culture, producing the cell suspension consisted of dividing cells in more than 80%. Harvested cells are then inoculated in plastic dishes, where the feeder layer have been seeded 40 hr before. The growth measurement is carried out with window counting method. The median generation time, thus obtained, is 24.5 hr and F value, an index of synchrony, is 0.4. The growth curve of this synchronous system is similar to the cumulative frequency distribution curve constructed with generation times of individual cells in a random culture which were determined through the cinematographic observation. The other growth parameters as G_1, S, G_2 and M of cells are determined to be 6.4 hr, 12.2 hr and 5.9 hr, respectively, by the autoradiography of cells which were pulsewise labeled with ^3H-thymidine. Those show a good agreement with the parameters found in randomly growing population. For reasons mentioned above, it would be concluded that the harvesting procedure assures a high degree of synchrony with this system and give a minimal disturbance on cell's physiology in the experimental condition.
千葉大学の論文
1966-03-28

FRUKTO細胞の増殖と同調培養法

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