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Fig. 1 | BMC Veterinary Research

Fig. 1

From: An EdU-based flow cytometry assay to evaluate chicken T lymphocyte proliferation

Fig. 1

Impact of cell culture medium on cell viability. Spleen mononuclear cells were isolated through a density gradient and cultured in RPMI-1640 supplemented with 5% fetal bovine serum (FBS), RPMI-1640 supplemented with 5% chicken serum (ChS), DMEM:F12 supplemented with 5% ChS, or FARMEM supplemented with 0.5% ChS. Cells were cultured in duplicate in the presence or absence of 1 μg/ml ConA. At 72 h post-plating, cell viability was evaluated using the 7AAD reagent. (a) Dot plots representing the flow cytometry analysis. (b) Comparison of cell viability in RPMI-1640 supplemented with 5% FBS and RPMI-1640 supplemented with 5% ChS. (c) Comparison of cell viability in DMEM:F12 supplemented with 5% ChS and FARMEM. (d) Biparametric FSC/SSC dot plot of stimulated cells with ConA in RPMI-1640 supplemented with 5% FBS, RPMI supplemented with 5% ChS, DMEM:F12 or FARMEM medium. Data are represented as the percentage of the 7AAD negative cells in relation to the total population. Results are expressed as the mean ± standard deviation. Significant differences are indicated by * (p = 0.0286). Each dot of the same color represents an independent experiment. In each experiment, the cells of 1 chicken were analyzed. Per sample, 30,000 events were acquired on a FACSMelody flow cytometer

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