CAJ | 학술논문

目的：探索H5N1亚型禽流感病毒在MDCK中增殖规律，确定最佳增殖条件。方法将H5N1亚型禽流感病毒接种到6孔板培养的MDCK细胞进行增殖试验，检测不同病毒感染量、不同浓度TPCK-胰酶，接毒后不同时间病毒的HA滴度。根据确定的最佳增殖条件将病毒接种到微载体培养的MDCK细胞中进行大规模增殖。结果：最佳病毒增殖条件接毒量MOI为5×10-4、TPCK-胰酶浓度为4μg/mL，在5 L生物反应器中重复验证，获得稳定的试验结果，病毒血凝价最高为8 log2。结论：本研究为禽流感疫苗的生物反应器规模化生产奠定了基础。
목적：탐색H5N1아형금류감병독재MDCK중증식규률，학정최가증식조건。방법장H5N1아형금류감병독접충도6공판배양적MDCK세포진행증식시험，검측불동병독감염량、불동농도TPCK-이매，접독후불동시간병독적HA적도。근거학정적최가증식조건장병독접충도미재체배양적MDCK세포중진행대규모증식。결과：최가병독증식조건접독량MOI위5×10-4、TPCK-이매농도위4μg/mL，재5 L생물반응기중중복험증，획득은정적시험결과，병독혈응개최고위8 log2。결론：본연구위금류감역묘적생물반응기규모화생산전정료기출。
To explore the regularity for the multiplication of avian influenza virus subtype H5N1 in MDCK cell culture system and determine the optimal proliferation conditions. H5N1 subtype of avian influenza virus was in-oculated into the MDCK cell growing on 6 well plate, and the HA titers of virus at different time were detected in the conditions of different infectious doses, and different concentrations of TPCK-trypsin. The optimal conditions were determined. Then the H5N1 subtype avian influenza virus was grown in microcarrier-based MDCK cell. It was demonstrated that high virus yield with a hemagglutination unit of 8 log2 (1:256) could be obtained under the optimal conditions of multiplication. The result indicated the H5N1 subtype avian influenza virus could be pro-duced in microcarrier-based MDCK cell in a large-scale culture system with a high virus yield and demonstrates the feasibility of the development of mammalian cell-based in influenza vaccine in microcarrier culture systems.