Peripheral blood is commonly sampled to assess the health status of human and veterinary patients. Venous blood collection is a minimally invasive procedure, and in the horse, the common collection site is the jugular vein. Post blood collection, sample processing for leukocyte enrichment can vary by research laboratory with the potential to yield different effects on the enriched cells and their function. The focus of the present study was to compare a common blood dilution-leukocyte enrichment technique using a Histopaque gradient medium (His) to a modified leukocyte buffy coat syringe-lymphocyte separation medium technique (Syr- LSM) with peripheral blood from 12 healthy horses. The endpoints examined included cell recovery/mL of blood, cell viability, leukocyte enrichment purity, leukocyte cell marker subset phenotype, leukocyte spontaneous and mitogen-induced proliferation and secretory TNFα concentrations. Leukocyte cell recovery/mL of whole blood and cell viability was significantly increased in enriched leukocytes from the Syr-LSM technique. Interestingly, the percentage of CD8+ and CD21+ were significantly increased with the His technique as was Con A-induced proliferation. Still, leukocyte cell purity and TNFα concentrations from the 72 h cell culture supernatants were comparable across the two enrichment techniques. To summarize, the type of whole blood leukocyte enrichment technique employed can affect the results of immunologic assay endpoints possibly altering data interpretation.

中文翻译：

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外周血富集技术的可变性可以改变马白细胞的细胞性，生存能力和功能。

通常采集外周血以评估人类和兽医患者的健康状况。静脉血液采集是一种微创手术，在马中，常见的采集部位是颈静脉。采血后，研究实验室对白细胞富集的样品处理方法可能有所不同，可能会对富集的细胞及其功能产生不同的影响。本研究的重点是比较使用12个健康马的外周血用组织细胞梯度梯度培养基（His）与改良的白细胞血沉棕黄层注射器-淋巴细胞分离培养基技术（Syr- LSM）进行的普通血液稀释-白细胞富集技术。检查的终点包括细胞回收率/血液mL，细胞活力，白细胞富集纯度，白细胞标志物亚型表型，白细胞自发和有丝分裂原诱导的增殖和分泌性TNFα浓度。通过Syr-LSM技术富集的白细胞中白细胞回收率/全血和细胞活力的mL显着提高。有趣的是，通过His技术，CD8 +和CD21 +的百分比显着增加，Con A诱导的增殖也是如此。尽管如此，在两种富集技术中，来自72 h细胞培养上清液的白细胞纯度和TNFα浓度是可比的。总而言之，采用的全血白细胞富集技术的类型可能会影响免疫测定终点的结果，可能会改变数据解释。通过Syr-LSM技术富集的白细胞中白细胞回收率/全血和细胞活力的mL显着提高。有趣的是，通过His技术，CD8 +和CD21 +的百分比显着增加，Con A诱导的增殖也是如此。尽管如此，在两种富集技术中，来自72 h细胞培养上清液的白细胞纯度和TNFα浓度是可比的。总而言之，采用的全血白细胞富集技术的类型可能会影响免疫测定终点的结果，可能会改变数据解释。通过Syr-LSM技术富集的白细胞中白细胞回收率/全血和细胞活力的mL显着提高。有趣的是，通过His技术，CD8 +和CD21 +的百分比显着增加，Con A诱导的增殖也是如此。尽管如此，在两种富集技术中，来自72 h细胞培养上清液的白细胞纯度和TNFα浓度是可比的。总而言之，采用的全血白细胞富集技术的类型可能会影响免疫测定终点的结果，可能会改变数据解释。在两种富集技术中，来自72 h细胞培养上清液的白细胞纯度和TNFα浓度可比。总而言之，采用的全血白细胞富集技术的类型可能会影响免疫测定终点的结果，可能会改变数据解释。在两种富集技术中，来自72 h细胞培养上清液的白细胞纯度和TNFα浓度可比。总而言之，采用的全血白细胞富集技术的类型可能会影响免疫测定终点的结果，可能会改变数据解释。