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of any two were tested. Only 10% alligator serum and 5% alligator-5%
fetal calf serum supported in mitogen stimulation of lymphocytes. Al
though significant stimulation was obtained in cultures supplemented
with an equimixture of alligator and fetal calf sera, stimulation in
dices were lower than those obtained from 10% alligator serum supple
mented cultures and therefore 10% alligator serum was used routinely.
Not all alligator sera were supportive as a supplement and it was
necessary to test new alligator supplement sources in mitogenic assays
to determine their suitability. Tests of eight serum pools (> 10.
individual bleedings per pool) obtained from 2-3 yr old alligators
and four individual alligators > 10 yr old are presented in Table 13.
Individual sera from older alligators were more effective than pools of
sera from younger alligators. Since large volumes (200-500 ml) could
be obtained from individual bleedings of 100-225 kg alligators, the
older alligators were used exclusively as sources of serum in subse
quent experiments.
Although statistically significant stimulation of alligator lym
phocytes cultured with mitogens was obtained using 10% alligator serum
supplemented MEM (0.117 M NaCl), severe cell clumping and loss of
viability were noted when cells were suspended in the culture medium.
To determine if the salt concentrations in the medium were appropriate
ly matched to alligator serum levels, three alligator sera (obtained from
individual bleedings) were analyzed by the Blood Chemistry Lab (Depart
ment of Pathology, J. Hillis Miller Health Center). Comparisons of the
chemistry lab reports with the GIBCO MEM formulations revealed a repro
ducible difference in the NaCl concentrations. On the basis of this
finding an experiment in which alligator peripheral blood lymphocytes