CAJ | 학술논문

目的：轮状病毒基因重配株Ls（ G3型）在生物反应器微载体培养Vero细胞条件的优化。方法采用3 L生物反应器微载体培养Vero细胞，观察Ls 株在不同病毒感染复数（0．001、0．002、0．010、0．040 MOI）、不同温度（34．5℃和35．5℃）、不同病毒收获时间（24和48 h）对病毒增殖的影响。根据病毒滴度和收获量筛选出最适MOI、培养温度及病毒的收获时间。结果以0．002 MOI接种Vero细胞，温度为34．5℃培养病毒，滴度最高达7．50 lgCCID50／mL；48 h可连续收获4次病毒液，且收获总量及病毒滴度均高于24 h。结论通过对Ls株在生物反应器微载体Vero细胞培养条件的优化，获得的病毒液滴度高及连续培养多次收获量增加的有效方法，为进一步规模化培养奠定了基础。
목적：륜상병독기인중배주Ls（ G3형）재생물반응기미재체배양Vero세포조건적우화。방법채용3 L생물반응기미재체배양Vero세포，관찰Ls 주재불동병독감염복수（0．001、0．002、0．010、0．040 MOI）、불동온도（34．5℃화35．5℃）、불동병독수획시간（24화48 h）대병독증식적영향。근거병독적도화수획량사선출최괄MOI、배양온도급병독적수획시간。결과이0．002 MOI접충Vero세포，온도위34．5℃배양병독，적도최고체7．50 lgCCID50／mL；48 h가련속수획4차병독액，차수획총량급병독적도균고우24 h。결론통과대Ls주재생물반응기미재체Vero세포배양조건적우화，획득적병독액적도고급련속배양다차수획량증가적유효방법，위진일보규모화배양전정료기출。
Objective To optimize the key condition for culture of rotavirus reassortant strain Ls( Type 4) by using vero cells adsorpted on microcarriers cultured in bioreactor.Methods The effect to multiplication of the virus was investigated, Vero cells cultured in 3 L bioreactor as cell matrix, then inoculated with Ls strain at MOIs of 0.001, 0.002, 0.010 and 0.040, in different temperatures(34.5℃or 35.5℃) and different harvest intervals(24 h or 48 h), on which the optimal MOI, temperature and havest interval were decided.Results Under the condition of 0.002 MOI, 34.5 ℃, Ls was cul-tured well with a titer up to 7.50 lgCCID50/mL, and a continuous four harvests obtained in 48 h, and the total harvest and the virus titers were higher than cultured in 24 h.Conclusion An effective method for increasing rotavirus titer was devel-oped by optimizing the condition for culture of Ls on Vero cells in biorector,on which the foundation was developed in a large scale preparation of rotavirus vaccine, live.