Faculty of Veterinary Science, Department of Clinical Sciences and Public Health, Mahidol University, Salaya, Phutthamonthon, Nakhon Pathom, Thailand.
Faculty of Veterinary Science, Department of Pre-clinic and Applied Animal Science, Mahidol University, Salaya, Phutthamonthon, Nakhon Pathom, Thailand.
PLoS One. 2022 Jul 22;17(7):e0269108. doi: 10.1371/journal.pone.0269108. eCollection 2022.
The objectives of this study were to evaluate the proper anticoagulants coated in blood-collecting tube for the peripheral blood mononuclear cells (PBMCs) isolation and to evaluate the proper culture temperature for the Varanus salvator's PBMCs, in addition, the hematological characteristics also reported. The heparin treated blood (n = 10) and EDTA treated blood (n = 10) from Varanus salvator were obtained for PBMCs evaluation. The PBMCs obtained from the heparin treated blood was significantly higher than that of EDTA treated blood during the culture period (P < 0.05) indicated heparin would be more appropriated anticoagulant for blood collection. The PBMCs cultured under 37°C and 27°C were not significantly difference on first three days but 37°C showed significantly higher effect in the following days (P < 0.05) indicated both temperatures can be used which 37°C should be an optimal for PBMCs preparation. The peripheral blood cells of Varanus salvator (n = 49) were analyzed for hematological profiles and characteristics which the number of erythrocytes 1.19 ± 0.04 x 1012/L (1.17-1.35 x 1012/L) and WBC 2.41 ± 0.13 x 109/L (2.29-2.81 x 109/L) with absolute differential count of heterophils 0.92 ± 0.02 x 109/L (0.87-0.95 x 109/L), lymphocytes 1.17 ± 0.01 x 109/L (1.15-1.23 x 109/L), azurophils 0.40 ± 0.01 x 109/L (0.37-0.42 x 109/L), basophils 0.000 ± 0.001 x 109/L (0.004-0.011 x 109/L) and monocytes 0.027 ± 0.002 x 109/L (0.028-0.039 x 109/L). These results would play an important role on the cell immunological studies of the Varanus salvator in the future.
本研究的目的是评估适合用于外周血单个核细胞（PBMCs）分离的涂有抗凝剂的采血管，并评估培养巨蜥 PBMCs 的合适温度，此外还报告了血液学特征。从巨蜥 salvator 中获得肝素处理的血液（n = 10）和 EDTA 处理的血液（n = 10）用于 PBMCs 评估。在培养期间，肝素处理的血液中获得的 PBMCs 明显高于 EDTA 处理的血液（P < 0.05），表明肝素是更适合采血的抗凝剂。在 37°C 和 27°C 下培养的 PBMCs 在最初三天没有显著差异，但 37°C 在随后几天显示出显著更高的效果（P < 0.05），表明两种温度都可以使用，其中 37°C 应为 PBMCs 制备的最佳温度。分析了 49 只巨蜥 salvator 的外周血细胞的血液学特征，红细胞数为 1.19 ± 0.04 x 1012/L（1.17-1.35 x 1012/L），白细胞数为 2.41 ± 0.13 x 109/L（2.29-2.81 x 109/L），嗜中性粒细胞绝对差异计数为 0.92 ± 0.02 x 109/L（0.87-0.95 x 109/L），淋巴细胞 1.17 ± 0.01 x 109/L（1.15-1.23 x 109/L），嗜酸性粒细胞 0.40 ± 0.01 x 109/L（0.37-0.42 x 109/L），嗜碱性粒细胞 0.000 ± 0.001 x 109/L（0.004-0.011 x 109/L）和单核细胞 0.027 ± 0.002 x 109/L（0.028-0.039 x 109/L）。这些结果将在未来对巨蜥 salvator 的细胞免疫学研究中发挥重要作用。
