Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Groennegaardsvej 15, DK-1870 Frederiksberg C, Denmark.
Novo Nordisk A/S, Novo Nordisk Park, DK-2760, Måløv, Denmark.
J Pharmacol Toxicol Methods. 2020 May-Jun;103:106680. doi: 10.1016/j.vascn.2020.106680. Epub 2020 Feb 11.
Repeated blood sampling is a common procedure in laboratory mice, but at present it is unknown which technique has the least impact on the animals when large or repeated blood samples are required. Retro-bulbar sinus puncture is a reliable technique but has been shown to cause many changes in the animals, why sublingual and facial vein puncture have been suggested as suitable alternatives. This study investigated 1) which of the three blood sampling techniques had the least impact on nest building activity, level of faecal corticosterone metabolites, body weight, fur status, and macroscopic changes, 2) whether the blood sampling techniques gave rise to variation in blood quality between blood samples, and 3) whether sublingual and facial vein puncture should be performed with or without anaesthesia in female C57BL/6 mice.
Three hundred and sixty C57BL/6 female mice divided into five batches were included in the study and randomized to a short (blood sampling on Day 8, 9 and 10) or a long protocol (blood sampling on Day 8, 15 and 22). Each protocol consisted of six identical groups: sublingual vein puncture (SVP), sublingual vein puncture in isoflurane (SVPiso), facial vein puncture (FVP), facial vein puncture in isoflurane (FVPiso), retro-bulbar sinus puncture (RBP), and a control group (CONTROL) with only scruffing being performed. At baseline (Day 2) nest building activity (NBA) was assessed and faecal pellets collected for evaluation of faecal corticosterone metabolites (FCM). The day after each blood sampling day NBA and FCM were reassessed.
None of the blood sampling techniques proved to be superior to the others in any of the measured parameters. Finally, sublingual and facial vein puncture performed under anaesthesia gave rise to variation in the quality of the blood. A refinement of all three techniques are therefore warranted.
在实验室小鼠中，反复采血是一项常见的操作，但目前尚不清楚在需要大量或反复采血时，哪种技术对动物的影响最小。球后窦穿刺是一种可靠的技术，但已证明它会引起动物的许多变化，因此建议使用舌下和面部静脉穿刺作为合适的替代方法。本研究调查了 1）三种采血技术中哪一种对筑巢活动、粪便皮质酮代谢物水平、体重、皮毛状况和宏观变化的影响最小，2）采血技术是否会导致血液质量在血液样本之间发生变化，3）在 C57BL/6 雌性小鼠中，是否应在麻醉下或不麻醉下进行舌下和面部静脉穿刺。
将 360 只 C57BL/6 雌性小鼠分为 5 批纳入本研究，并随机分为短期（第 8、9 和 10 天采血）或长期方案（第 8、15 和 22 天采血）。每个方案包括 6 个相同的组：舌下静脉穿刺（SVP）、异氟烷麻醉下舌下静脉穿刺（SVPiso）、面部静脉穿刺（FVP）、异氟烷麻醉下面部静脉穿刺（FVPiso）、球后窦穿刺（RBP）和仅进行抓痒的对照组（CONTROL）。在基线（第 2 天）评估筑巢活动（NBA）并收集粪便样本来评估粪便皮质酮代谢物（FCM）。在每次采血后的第二天重新评估 NBA 和 FCM。
在任何测量参数中，没有一种采血技术被证明优于其他技术。最后，在麻醉下进行的舌下和面部静脉穿刺会导致血液质量的变化。因此，所有三种技术都需要进一步改进。
