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Thesis Advisor李佐虎
Degree Grantor中国科学院研究生院
Other AbstractInsect cell culture was used to produce wild baculovirus and pro-uerokinase (pro-UK) in this study. The main research was divided into 5 parts: 1 Insect cell growth was studied for serum was verified through the test of inoculum effect and conditional medium. Sf9 cells was adapted in 2% serum medium. Trypotse phosphate broth (TPB) was added as a serum substitute. Compared to 10% serum medium, adapted cells obtained similar growth in 2% serum medium with 2% TPB supplemented, while the culture cost reduced 50%. 2 Sf9 cell can grow in various culture vessels. 50-100 rpm was suitable for suspension culture. For batch culture, the maximum cell density was 18.6 * 10~5/ml in spinner flask, 17.5 * 10~5/ml in 2-L agitator bioreactor, 23.5 * 10~5/ml in air-liquid-lift bioreactor. Cells raised to 23.8 * 10~5/ml in half medium replacement and to 25.4 * 10~5/ml in TPB feeding culture. 3 Effects of multiplicity of infection (MOI) was dependent on time of infection. Low MOI required early infection and long culture time, while high MOI was reverse. Pro-UK yields were 420 IU/ml in spinner flask and 530 IU/ml in 2-L agitator bioreactor with MOI=3. Medium replacement and fed-batch can enhanced baculovirus and pro-UK yields. The pro-UK yield reached 820 IU/ml in 1% serum medium with 2% TPB supplemented. 4 Temperature oscillation was introduced to insect cell-baculovirus culture system. The optimum temperature oscillation in 12 hr for baculovirus propagation was 24-28 ℃. Compared to 28 ℃ culture, the baculovirus yields raised about 30%. 5 Phenomenological model of cell growth was established on experimental work. This model could describe the whole process of insect cell baculovirus culture system and coincide well with the experimental result.
Document Type学位论文
Recommended Citation
GB/T 7714
常韶华. 昆虫细胞-杆状病毒培养体系的研究[D]. 中国科学院研究生院,1998.
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