PK-15细胞在微载体悬浮放大培养及PCV2增殖工艺研究

发布时间：2018-04-20 09:21

本文选题：PK-细胞 + 微载体　；参考：《西南农业学报》2017年02期

【摘要】：为建立在生物反应器内微载体逐级放大培养PK-15细胞和增殖PCV2技术。采用分批式培养方法,研究了PK-15细胞在微载体胰酶消化放大悬浮培养及猪圆环病毒2型(PCV2)增殖工艺。结果表明,5 g/L的微载体和高糖DMEM下,采用4.0×105cells/m L的初始接种密度及培养至第2天进行换液操作工艺可以获得最佳的PK-15细胞生长效能。胰酶消化转移将PK-15细胞从1 L反应器放大至3 L反应器,微载体上细胞贴附均匀、生长旺盛,培养3 d细胞密度可达34.3×105cells/m L。采用感染复数为0.1的接毒比例,细胞接毒后在微载体上传至第3代收获毒液可获得最高的PCV2增殖效价107.25TCID50/m L。为PCV2疫苗的生物反应器规模化生产奠定了基础。
[Abstract]:In order to establish the technique of PK-15 cell culture and PCV2 proliferation in bioreactor, the microcarriers were amplified step by step. Batch culture method was used to study the process of PK-15 cell proliferation in microcarrier trypsin digestion and suspension culture and porcine circovirus type 2 (Porcine circovirus 2) proliferation. The results showed that under 5 g / L microcarrier and high glucose DMEM, the optimal growth efficiency of PK-15 cells could be obtained by using the initial inoculation density of 4.0 脳 105cells/m L and the operation of changing solution to the second day. The PK-15 cells were amplified from 1 L reactor to 3 L reactor by trypsin digestion and metastasis. The cells on the microcarriers were uniformly attached and grew vigorously. The cell density reached 34.3 脳 105cells/m / L after 3 days of culture. The highest PCV2 proliferation titer (107.25TCID50/m L) was obtained by inoculating the cells with the infected complex number of 0.1, and then uploading it to the third generation harvest venom after inoculating with the microcarrier. It lays a foundation for the bioreactor production of PCV2 vaccine.
【作者单位】：江苏省农业科学院国家兽用生物制品工程技术研究中心农业部兽用生物制品工程技术重点实验室;江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：江苏省农业自主创新项目[CX(14)5044] 农业部公益性行业(农业)科研专项经费项目(201303046) 江苏省农业支撑计划(BE2012370)
【分类号】：S852.651

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