中文摘要：

【目的】建立一种适用于蜜蜂源的细胞培养方法,为蜜蜂源细胞培养和蜂病毒的研究奠定基础。【方法】比较在Grace和WH2两种昆虫细胞培养基中培养的中华蜜蜂Apis cerana cerana幼虫原代细胞状况,筛选出适用于中华蜜蜂幼虫原代细胞培养的最佳培养基,并通过细胞活力比较,确定用于中华蜜蜂幼虫原代细胞培养的适宜胎牛血清（fetal bovine serum,FBS）浓度。在此基础上,用该原代细胞接种中华蜜蜂囊状幼虫病毒（Chinese sacbrood bee disease,CSBV）,并通过实时定量RT-PCR方法对病毒复制情况进行检测。【结果】相对于WH2培养基,在Grace培养基中生长的细胞大而圆,透明,边缘整齐,无颗粒物,活力明显高于WH2培养,且含15%FBS的Grace培养基更适合于中华蜜蜂幼虫原代细胞培养。CSBV接种在该原代细胞,能够复制增殖,同时伴随宿主细胞快速分裂。【结论】中华蜜蜂幼虫原代细胞培养基在含15%FBS的Grace中能够良好生长,并且CSBV可以在该原代细胞中进行复制。

英文摘要：

[ Aim] To establish an applicable culture method for honeybee sourced cells and to lay the foundation for culture of honeybee cells and related virus research. [ Methods ] The premium culture medium and the corresponding fetal bovine serum （FBS） concentration were screened by comparing the growth status of the primary cells of the Chinese bee larvae cultured in Grace＇ s medium and WH2 medium, respectively. Then, the cultured primary cells were inoculated with Chinese acbrood virus （CSBV）, and the virus replication was detected with real-time quantitative RT-PCR. [ Results] Cells cultured in Grace＇ s medium were large and round, transparent, and regularly edged with no particulate matters, and had significant higher viability than those cultured in WH2 medium. The economical and applicable FBS concentration was determined to be 15% based on the comparison of the cell viability cultured in Grace＇ s medium with various concentrations of FBS. The inoculated CSBV virus could replicate and proliferate rapidly along with the division of the host cells. [ Conclusion] The primary cells of Chinese bee larvae grow well in Grace＇ s medium with 15% FBS, in which the replication of CSBV can proceed.