背角无齿蚌四种组织细胞原代培养的比较  

Comparison on Primary Culture of Four Tissular Cells in Anodonta woodiana

作　　者：阎明军 陈修报[2] 刘洪波[2] 杨健[1,2] YAN Ming-Jun;CHEN Xiu-Bao;LIU Hong-Bo;YANG Jian(Wuxi Fisheries College,Nanjing Agricultural University,Wuxi 214081;Laboratory of Fishery Microchemistry,Freshwater Fisheries Research Center,Chinese Academy of Fishery Sciences,Wuxi 214081,China)

机构地区：[1]南京农业大学无锡渔业学院,无锡214081 [2]中国水产科学研究院淡水渔业研究中心渔业微化学研究室,无锡214081

出　　处：《动物学杂志》2023年第5期751-759,共9页Chinese Journal of Zoology

基　　金：中国水产科学研究院基本科研业务费(No.2019GH10),中国水产科学研究院科技创新团队项目(No.2020TD18);国家自然科学基金青年科学基金项目(No.31502166)。

摘　　要：以背角无齿蚌(Anodonta woodiana)血细胞(含颗粒细胞和透明细胞)、消化腺细胞(含上皮样细胞和圆形小细胞)、外套膜细胞(含上皮样细胞、透明细胞、圆形大细胞和圆形小细胞)、斧足细胞(含上皮样细胞、透明细胞、圆形大细胞和圆形透亮细胞)为材料,比较了上述各类细胞的原代培养特征以及不同条件下的细胞存活率,优化了细胞制备及细胞活性鉴定的方法。结果显示,上述各类组织细胞存活率受不同培养温度、培养时间和培养基种类的影响,其中,血细胞96 h在20℃下、培养基2中的存活率最高,为94.67%±0.47%;消化腺细胞48 h在20℃下、培养基3中的存活率最高,为93.67%±1.70%;外套膜细胞48h在15℃下、培养基1中的存活率最高,为93.67%±1.7%;斧足细胞48h在15℃下、培养基1的存活率最高,为94.33%±0.94%。相较于25℃,在20℃和15℃下,4种组织细胞96h的存活率更为理想。本研究旨在为背角无齿蚌组织细胞系的建立及贝类细胞水平毒理学的研究提供基础数据。[Objectives]Animal cell culture technologies become more crucial in ecotoxicological researches.Due to some technical difficulties,studies on tissular cell culture of freshwater mussels are still limited.Although primary culture of gill cell in Anodonta woodiana was succeeded in previous study,breakthroughs have not yet been obtained for other tissues.Therefore,we intend to expand the primary culture of A.woodiana cells from four tissues(hemocyte,digestive gland,mantle,and foot).[Methods]The healthy 2-year-old“standardized”A.woodiana were used to obtain tissue cells.Different culture conditions(e.g.,culture media,temperature)were designed to comparatively reveal survival rates and cell viabilities of the above four tissular cells so as to optimize the culture conditions for the primary culture.[Results]The results showed that the survival rates of the above four tissular cells were affected by different culture temperature,time and medium type.The survival rates were 94.67%±0.47%in culture medium 2 at 20℃for 96 h for the hemocytes,93.67%±1.70%in medium 3 at 20℃for 48 h for the digestive gland cells,93.67%±1.70%in medium 1 at 15℃for 48 h for the mantle cells,and 94.33%±0.94%in medium 1 at 15℃for 48 h for the foot cells(Fig.2).Compared to 25℃,the survival rates of four tissular cells were higher at 20℃and 15℃for 96 h.[Conclusion]The study obtained the breakthrough on primary culture of hemocyte,digestive gland,mantle,and foot cells in A.woodiana.The study provides basic data for the establishment of cell lines of A.woodiana.It can also provide theoretical support for subsequent studies of toxicology at the cellular level of freshwater mussels in the future.

关 键 词：背角无齿蚌 组织细胞 温度 原代培养 存活率 

分 类 号：Q952[生物学—动物学]