Evaluation of chicken kidney and chicken embryo kidney cultures for the large-scale growth of attenuated influenza virus master strain A/Ann/Arbor/6/60-ca.
Evaluation of chicken kidney and chicken embryo kidney cultures for the large-scale growth of attenuated influenza virus master strain A/Ann/Arbor/6/60-ca.
复制标题
鸡肾和鸡胚肾培养物对减毒流感病毒主株 A/Ann/Arbor/6/60-ca 大规模生长的评价。
DOI：
10.1016/s0264-410x(85)90245-2
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发表时间：
1985
期刊：
影响因子：
5.5
通讯作者：
J. A. Paul
中科院分区：
文献类型：
--
作者：
G. Tannock;D. Bryce;J. A. Paul
研究了鸡原代肾（CK）和鸡胚肾（CEK）细胞作为冷适应流感减毒疫苗主株A/Ann Arbor/6/60 （A/AA/6/60-ca）生长的可能底物。在两种细胞类型中，每ml培养液的产量均为106 - 107tcid50。人二倍体菌株MRC-5的产量低约100倍。在4°C下使用过夜胰蛋白酶化步骤，从CEK细胞获得的培养物比CK细胞获得的培养物更具可重复性。在滚轮培养中生长的CEK细胞与在微载体表面生长的CEK细胞的每个胚胎的产量相当。这些产量低于从全胚尿囊液中获得的产量。在原代胰蛋白酶化、从培养的CK原代单层分散或在微载体上培养后，将CEK或CK细胞冷冻保存均不成功。在CEK培养物中生长后，a /AA/6/60-ca的冷适应表型得以保留，且在小鼠中检测到CEK病毒和尿囊生长病毒的免疫原性没有差异。A/AA/6/60的尿囊培养制剂的每感染单位蛋白浓度低于在CEK中培养的制剂
Primary chicken kidney (CK) and chicken embryo kidney (CEK) cells were evaluated as possible substrates for growth of the cold-adapted attenuated influenza vaccine master strain A/Ann Arbor/6/60 (A/AA/6/60-ca). Yields of 106–107TCID50per ml of culture fluid were obtained in either cell type. Yields from the human diploid strain MRC-5 were ∼100-fold less. More reproducible cultures were obtained from CEK cells, using an overnight trypsinization step at 4°C than from CK cells. Comparable yields per embryo were obtained from CEK cells grown in roller cultures to those grown on the surface of microcarriers. These yields were less than those obtained from the allantoic fluids of whole embryos. Frozen storage of CEK or CK cells, after primary trypsinization, dispersal from a cultured CK primary monolayer or culture on microcarriers, was unsuccessful The cold-adapted phenotype of A/AA/6/60-cawas retained after growth in CEK cultures and no differences in immunogenicity were detectable in mice between CEK- and allantoic-grown virus. Allantoic-grown preparations of A/AA/6/60-cacontained a lower protein concentration per infectious unit than those grown in CEK