低血清培养PK-15细胞及其增殖猪圆环病毒2型的研究

发布时间：2018-05-30 18:50

本文选题：PK-细胞 + 低血清培养基　；参考：《动物医学进展》2016年09期

【摘要】：依次采用含60、30、20、10mL/L血清浓度的低血清培养基驯化PK-15细胞,并给驯化好的细胞上接种猪圆环病毒2型(PCV-2),以确定PCV-2在该培养体系下的生长情况。结果用含60、30、20mL/L血清浓度的低血清培养基各进行3代次驯化,PK-15细胞能完全适应且生长状态良好;当血清浓度降至10mL/L时,传代1次细胞无法保持良好状态,细胞出现贴壁较差、生长停滞等现象。各血清浓度培养体系中细胞生长曲线测定结果显示,在细胞培养的0、24、48、72、96h各组细胞密度与常规培养的对照组差别不明显。因此用该低血清培养体系培养PK-15细胞时,血清最低添加量为20mL/L。用该体系培养的PK-15细胞接种PCV-2后,通过荧光抗体染色测定病毒滴度。结果显示,低血清培养的PCV-2病毒滴度为10~(6.5)TCID_(50)/mL,与常规条件培养的PCV-2对照(病毒滴度为10~(6.375 )TCID_(50)/mL)差别不明显,表明该体系可用于PCV-2的增殖。表明研究建立了PK-15细胞低血清培养PCV-2体系,为PCV-2的相关研究奠定了基础。
[Abstract]:The PK-15 cells were acclimated on a low serum medium containing 60 ~ 30 ~ 20ml / L serum concentration and inoculated with porcine circovirus type 2 PCV-2 to determine the growth of PCV-2 in this culture system. Results the PK-15 cells were acclimated completely and grew well when the serum concentration of PK-15 was reduced to 10mL/L, and the cells could not maintain a good condition when the serum concentration was reduced to 10mL/L, and the cells had poor adherence to the wall, and the cells could not be acclimated to the medium with low serum concentration of 60 ~ 30 ~ 20 mL / L for 3 passages. Stagnation of growth, etc. The results of cell growth curve in each serum concentration culture system showed that there was no significant difference in cell density between the control group and the normal culture group at 7296 h after cell culture. Therefore, when PK-15 cells were cultured in this low serum culture system, the minimum serum addition was 20ml / L. The PK-15 cells cultured in this system were inoculated with PCV-2 and the virus titers were detected by fluorescent antibody staining. The results showed that the titer of PCV-2 virus cultured in low serum was 10 ~ (6. 5) TCID / mL, which was not significantly different from that of PCV-2 cultured in normal condition (the virus titer was 10 ~ 6. 375 / mL), which indicated that the system could be used for the proliferation of PCV-2. The results showed that the PCV-2 system of PK-15 cells cultured with low serum was established, which laid a foundation for the study of PCV-2.
【作者单位】：吉林大学动物医学学院;山东绿都生物科技有限公司;
【基金】：山东省重点研发计划项目(2015GSF121027)
【分类号】：S852.651

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