Culture conditions for arresting and stimulating the proliferation of a rainbow trout fibroblast cell line,RTG-2 |
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Authors: | L E J Lee A Martinez N C Bols |
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Institution: | (1) Department of Biology, University of Waterloo, N2L 3G1 Waterloo, Ontario, Canada |
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Abstract: | Summary Conditions for arresting and stimulating the proliferation of the rainbow trout fibroblast cell line RTG-2 have been examined
and the time course of events after stimulation determined. Quiescent populations were achieved in two ways. Cultures grown
to confluency without a medium change for at least 7 d had fewer than 5% of the cells in S phase and few mitotic figures.
Cultures deprived of serum, which could be done for up to 3 d without a loss in cell number, also achieved quiescence. After
3 d without serum, less than 1% of cells were in S phase and mitotic figures were infrequent. Addition to these cultures of
fresh serum-containing medium brought about the synchronous entry of cells into S phase and mitosis. For cultures in which
either the medium had been changed after 7 d without a change or serum-containing medium had been added after 3 d of serum
deprivation, DNA synthesis increased after a lag period of 20 to 24 h, was pronounced between 30 and 45 h, and then declined.
This was followed by a peak in the mitotic index. These protocols for arresting and subsequently stimulating RTG-2 proliferation
should allow the G1-S transition to be studied in a representative of teleosts.
This research was supported by Natural Sciences and Engineering Research Council of Canada grant to N. C. B. |
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Keywords: | trout fibroblasts cell proliferation synchronization |
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