Jo Eun Jung, Bae Eunjin, Yoon Jeong-Hwan, Kim Ji Yeon, Han Jin Soo
Department of Laboratory Animal Medicine, College of Veterinary Medicine, Konkuk University, Seoul, 05029, Republic of Korea.
Biomedical Research Institute, Kyungpook National University Hospital, Daegu, 41944, Republic of Korea.
Lab Anim Res. 2021 May 6;37(1):12. doi: 10.1186/s42826-021-00090-4.
Blood collection is an important procedure used in animal experiments. Blood collection methods that reduce pain, injury, and stress in experimental animals are important with regard to animal ethics. Various comparative studies of blood collection methods have been reported; however, there are no comparative studies on serial blood collection considering animal ethics. To suggest simple methods that minimize pain during serial blood collection, we compared the retroorbital plexus (RP) and facial vein (FV) blood collection methods performed by both experienced and novice groups. The experienced and novice groups collected up to 0.4 mL of blood via the RP and FV methods every second day for 2 weeks. After blood collection, all mice were evaluated by corticosterone concentrations for stress, hematological, immunological, and histological analyses.
We found that the FV methods reduced the collection time, pain, distress, tissue damage and lasting harms without anesthesia. Corticosterone concentrations in the peripheral blood were decreased in mice subjected FV methods compare with those subjected to RP methods. The proportion of granulocytes and monocytes, such as macrophages in the peripheral blood and spleen, was decreased in mice subjected to FV methods compared with that in mice subjected to RP methods in both experienced and novice groups. White blood cells were infiltrated in RP areas with severe tissue damage and inflammation.
With respect to animal ethics, we suggest that the FV method, a simple and fast technique that can easily be performed by both experienced and novice researchers, is suitable for serial blood collection.
采血是动物实验中的一项重要操作。在动物实验中，能减轻实验动物疼痛、损伤和应激的采血方法对于动物伦理而言至关重要。已有各种关于采血方法的比较研究报道；然而，尚无考虑动物伦理的连续采血比较研究。为了提出能将连续采血过程中的疼痛降至最低的简便方法，我们比较了经验丰富组和新手组采用眶后静脉丛（RP）和面部静脉（FV）采血的方法。经验丰富组和新手组每隔一天通过RP和FV方法采集多达0.4毫升血液，持续2周。采血后，对所有小鼠进行皮质酮浓度检测，以进行应激、血液学、免疫学和组织学分析。
我们发现FV方法在无需麻醉的情况下缩短了采血时间、减轻了疼痛、痛苦、组织损伤和持久伤害。与采用RP方法的小鼠相比，采用FV方法的小鼠外周血中皮质酮浓度降低。在经验丰富组和新手组中，与采用RP方法的小鼠相比，采用FV方法的小鼠外周血和脾脏中的粒细胞和单核细胞（如巨噬细胞）比例降低。RP区域出现白细胞浸润，伴有严重的组织损伤和炎症。
从动物伦理角度出发，我们建议FV方法是一种简单快速的技术，经验丰富和新手研究人员都能轻松操作，适用于连续采血。
