| Abstract |
Two continuous cell lines, designated AGE and AGF, were developed from
embryos of the ebony strain of the cotton boll weevil. Cultures were
initiated from minced tissues of several embryonic stages, using four
different media (Grace's, M & M, M199 and Schneider's, each supplemented
with 10% heat inacti vated fetal bovine serum and antibiotics). The most
successful primary cultures were derived from 24-30 hours old embryos on
Grace's and Schneider's media. Three weeks after initiation, one of the
cultures (AGE, started with 12 one day old embryos on Grace's medium)
showed extensive epithelial-like cell proliferation. The first passage
was obtained at week 4th and since then it has been subcultured 18 times
at 3-7 days intervals. Cells have been stored at -80?C using 7% Dimethyl
Sulfoxide as a cryoprotectant and revived successfully (90-95% recovery).
Karyotypic analysis at passage 7 showed high levels of ploidy. A second
culture (AGF, started with 12 two days old embryos on Grace's) showed
fibroblast like cell proliferation. The growth rate of this culture was
very slow during the first three passages (30-40 days intervals), but has
become notably faster during the last two (7-10 days). Once established,
AGE and AGF grew better on Schneider's.
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