Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations
Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations
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DOI：
10.1111/trf.13602
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发表时间：
2016-06-01
期刊：
影响因子：
2.9
通讯作者：
Bakchoul, Tamam
中科院分区：
文献类型：
--
作者：
Fuhrmann, Julia;Jouni, Rabie;Bakchoul, Tamam
背景NOD/SCID小鼠模型是研究人血小板(PLT)在体内存活的一种独特而先进的方法。与此同时，几个研究小组采用了这个模型来分析广泛的PLT抗体。研究小组之间在注射PLT的方法、抗体注射的数量和途径以及从动物身上采集的血液样本的制备方面存在差异，因此很难比较研究结果。研究设计和方法我们比较了通过尾静脉和眶后神经丛将人PLT注入NOD/SCID小鼠的存活率。用流式细胞仪检测小鼠外周血中人血小板的百分比。用两种不同的方法制备小鼠血液样本：1)直接固定全血样本和2)密度梯度离心法分离PLT。结果尾静脉注射人PLT后的回收率与经眼眶后注射相当(占循环PLT的13%比11%，P=0.401)。然而，尾静脉注射的PLT的存活率高于经眼眶注射的PLT(5小时后PLT的中位数存活率分别为84%和56%，P=0.025)。此外，与密度梯度离心法相比，直接固定的小鼠全血标本中循环人PLT的测定具有更好的重复性。结论NOD/SCID小鼠经尾静脉注射人PLT比经眶后注射人PLT的存活率更高。与密度梯度离心法相比，全血样本直接固定的结果具有更好的重复性。
BACKGROUNDThe NOD/SCID mouse model is a unique and sophisticated method to study the survival of human platelets (PLTs) in vivo. Meanwhile, several research groups adopted this model to analyze a wide range of PLT antibodies. Differences exist between the research groups regarding the method of PLT injection, the amount and route of antibody injection, and the preparation of blood samples collected from the animal, making it difficult to compare results between studies.STUDY DESIGN AND METHODSWe compared the survival of human PLTs infused into NOD/SCID mice via the tail vein or the retro-orbital plexus. The percentage of circulating human PLTs in the mouse circulation was determined by flow cytometry. Murine blood samples were prepared using two different methods: 1) direct fixation of whole blood samples and 2) isolation of PLTs by density gradient centrifugation.RESULTSRecovery of human PLTs after tail vein injection was comparable to retro-orbital injection (13% vs. 11% of all circulating PLTs, p=0.401). However, the survival rate of tail vein-infused PLTs was higher than that of retro-orbitally injected PLTs (median PLT survival after 5hr 84% vs. 56%, p=0.025). Moreover, we observed that determination of circulating human PLTs in directly fixed murine whole blood samples shows better reproducibility compared to the density gradient centrifugation method.CONCLUSIONSTail vein injection of human PLTs into the NOD/SCID mice is superior to retro-orbital injection in terms of human PLT survival. Direct fixation of whole blood samples allows better reproducibility of results compared to the density gradient centrifugation method.