文章摘要
樊廷俊.牙鲆鳃细胞与中国对虾淋巴细胞杂交细胞的体外培养[J].水产学报,2001,25(1):
牙鲆鳃细胞与中国对虾淋巴细胞杂交细胞的体外培养
In vitro culture of hybrid cells derived from flounder gill cells and shrimp lymphocytes
投稿时间:2014-04-16  修订日期:2014-04-16
DOI:
中文关键词: 牙鲆 中国对虾 杂交细胞 体外培养 连续性细胞系 细胞融合 病毒病
英文关键词: Paralichthys olivaceus  Penaeus chinensis  hybrid cell  in vitro culture
基金项目:国家海洋863 攻关项目(819-04-04)
作者单位地址
樊廷俊 青岛海洋大学海洋生命学院!山东青岛266003 青岛海洋大学海洋生命学院!山东青岛266003
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中文摘要:
      利用PCG介导的细胞融合方法,将已成系的牙鲆鳃细胞与中国对虾淋马细胞进行细胞杂交,对所获得的杂交细胞用MEM:MPS(1:1)混合培养液进行体外培养,培养5天后发现,杂交细胞长成了单层,传代培养后仍可继续生长和分裂,通过逐渐提高混合培养液中MPS的所占比例所筛选出的杂交细胞,目前已传12代,生长和分裂势头仍然十分旺盛,通过接种对虾杆现毒发现,10天后杂交细胞出现了明显的病变特征(细胞收缩变团圆,脱落,死亡),至14天细胞使几乎完全脱臂死亡,取病变细胞的培养上清0.3mL再加入到一瓶已长成单层的杂交细胞中,结果5天后杂交细胞也出出了明显的病变特和下,而未接种病毒的对照组杂交细胞生长正常,可禄步认为我们所得到的杂交细胞确为对虾(杆状)病毒的靶细胞。
英文摘要:
      The hybrid cells produced by PEG2mediated cell fusion method with gill cells from flounder , Paralichthys olivaceus , and lymphocytes from shrimp , Penaeus chinensis , were cultured in mixed medium of MEM∶MPS (1∶1) . After 5 days culture and observation , it was found that the hybrid cells cultured in the mixed medium of MEM∶MPS (1∶1) grew into monolayer and with good growing and proliferating appearance. As the concentration of MPS in the mixed medium was raised gradually, ideal hybrid cells had been screened out. By now , the obtained hybrid cells have been subcultured for 12 generations and still had very good growing and dividing appearance. Hybridoma cells acquired obvious disease appearance such as ,the cell contracted to round , disattachmentanddeath,afterinoculationwithwhitespotsyndromevirus (WSSV) ofshrimpfor 10 days, and died almost completely at the 14th day after inoculation. After 0. 3 mL culture supernatants of the cells with obvious disease appearance added into a new monolayer of normal hybrid cells , the hybrid cells acquired obvious disease appearance 5 days later , while the control cells still with normal appearance. It can be concluded that the hybrid cells obtained in this study are propably the target cells of shrimp viruses.
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