三种动物胰腺组织单细胞悬液制备效果比较  

Comparison on the Preparation of Pancreatic Tissue Single Cell Suspension from Three Kinds of Animals

作　　者：牛苗苗 刘军[1] 高伟[1] 曹江北[1] 陈华[1] NIU Miaomiao;LIU Jun;GAO Wei;CAO Jiangbei;CHEN Hua(the PLA General Hospital,Beijing 100853,China)

机构地区：[1]中国人民解放军总医院,北京100853

出　　处：《实验动物科学》2023年第5期38-42,共5页Laboratory Animal Science

基　　金：军队实验动物专项科研课题(SYDW[2020]01,SYDW[2020]02,SYDW[2020]03)。

摘　　要：目的比较巴马小型猪、SD大鼠、C57BL/6小鼠三种动物胰腺组织单细胞解离的效率和效果。方法分别将巴马小型猪、SD大鼠、C57BL/6小鼠麻醉后取新鲜胰腺组织,进行单细胞解离制备单细胞悬液。然后采用吖啶橙/碘化丙啶染色分别标记所有细胞核酸及死细胞核酸,应用全自动细胞荧光分析仪采集图像,观察细胞形态,计算细胞平均直径及平均圆度;根据吖啶橙/碘化丙啶染色情况,计算细胞活率、总细胞浓度和活细胞浓度。并对三种动物胰腺组织单细胞悬液各参数进行比较。结果三种动物胰腺组织单细胞悬液细胞形态完整、边界清楚,均有一定程度的结团。SD大鼠胰腺细胞的平均直径最大,C57BL/6小鼠细胞平均圆度最大。巴马小型猪和SD大鼠细胞活率较高,巴马小型猪总细胞浓度和活细胞浓度最高。结论成功制备了巴马小型猪、SD大鼠、C57BL/6小鼠三种动物胰腺组织单细胞悬液,为后续单细胞测序的开展奠定了基础。Objective To compare the efficiency and effect of single cell dissociation in pancreas tissue of Bama miniature pig,SD rat and C57BL/6 mouse.Method Fresh pancreatic tissue was taken from Bama miniature pig,SD rat and C57BL/6 mouse after anesthesia,and single cell suspension was prepared by single cell dissociation,respectively.Then,all cell nucleic acids and dead cell nucleic acids were labeled with acridine orange/propidium iodide staining,and the images were collected with automatic cell fluorescence analyzer to observe the cell morphology,calculate the average diameter and average roundness of cells.According to acridine orange/propidium iodide staining,cell viability,total cell concentration and living cell concentration were calculated.The parameters of pancreatic tissue single cell suspension from three kinds of animals were compared.Result The cells in the pancreas tissue single cell suspension from three kinds of animals,had complete morphology,clear boundary and certain degree of aggregation.The average diameter of pancreatic cells in SD rats is the largest,and the average roundness of cells in C57BL/6 mice is the largest.The cell viability of Bama minipigs and SD rats is higher,and the total cell concentration and living cell concentration of Bama minipigs are the highest.Conclusion The single cell suspension of pancreatic tissue from Bama miniature pig,SD rat and C57BL/6 mouse were successfully prepared,which laid a good foundation for subsequent single-cell sequencing.

关 键 词：巴马小型猪 SD大鼠 C57BL/6小鼠 胰腺 单细胞悬液 

分 类 号：Q95-3[生物学—动物学]