Nakata Makoto, Itou Takuya, Sakai Takeo
Nihon University Veterinary Research Center, Kanagawa, Japan.
J Vet Med Sci. 2007 Dec;69(12):1321-4. doi: 10.1292/jvms.69.1321.
Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.
通过密度梯度离心法从雪貂全血中分离出多形核细胞(PMNs)和外周血单核细胞(PBMCs)。使用50%的Percoll溶液(密度 = 1.066),离心后成功分离出PMNs和PBMCs;PMNs和PBMCs的纯度分别为94.2%和95.6%。为了评估分离出的雪貂PMNs的功能,我们用MCLA依赖性化学发光法测量了超氧化物的产生。分离出的雪貂PMNs对佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯(PMA)的反应动力学与人类PMNs相似。与能快速反应的人类PMNs不同,雪貂PMNs对N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸(fMLF)无反应。因此,作者建立了一种从雪貂全血中快速分离高纯度功能性PMNs群体的方法。