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题名	sf9 昆虫细胞悬浮培养的研究
作者	孙洪亮
学位类别	硕士
答辩日期	1998-06-01
授予单位	中国科学院研究生院
导师	李佐虎
中文摘要	可用细胞的比生长速率和葡萄糖、乳酸的代谢特征来表征昆虫细胞的状态。良好的细胞状态是尿激酶原高效表达的基础。建立了细胞培养动力学唯象模型。采用微生物发酵反应器成功地深层通气培养sf9昆虫细胞。深层通气大通气量时，细胞的代谢特征表明细胞对营养的需求可能改变。在已有的深层通气死亡速率模型的基础上建立了满足细胞对氧需求前提下的深层通气细胞死亡速率模型，把供氧和细胞损伤综合在一起考虑。本论文研究中发现昆虫细胞的比生长速率存在着锁定现象，锁定同细胞间的通讯有关。实验结果表明压力振荡对sf9昆虫细胞生长和产物表达没有显著性影响，但实验中没有进行全面和深入的研究，压力振荡对细胞生长和产物表达的影响值得进一步的研究。
索取号	昆虫细胞培养 ; 悬浮培养 ; 深层通气 ; 压力振荡
英文摘要	Insect cell-baculovirus culture can be used to produce many biologicals such as biopesticide and therapeutical proteins. As a vector for foreign gene, baculovirus has the following advantages: high expression level, post-translation modification and insertion of large foreign gene. Therefore, many researchers focus their attention on this expression system which has a good commercial prospect. The following studies about the insect cell-baculovirus expression system were done: 1. In batch culture with TC-100 media (10% serum and 2.5g/l TPB), the specific growth rate was 0.023±0.003h~(-1) (95% confidence) and the specific glucose consumption rate was 13±2μg/10~6 cells·h. The cell density 2.0×10~6 cells/ml was obtained in the same media. Normally, the specific glucose consumption rate decreased with growth slowing before the maximum cell density and decreased to 0 or very low level after the maximum cell density. In normal culture, Cell growth stopped with the depletion of glucose. Lactate slightly or hardly accumulated and was consumed after the middle of culture. The abnormal growth of sf9 insect cells was usually accompanied with the abnormal lactate accumulation and the decrease of the energy metabolic efficiency. According to the characteristics of sf9 insect cell growth and glucose consumption, the phenomenological mathematical model was suggested and fitted the experiments very well. 2. In sparging culture, even if cell growth was interrupted by four hours power failure, the cell growth and glucose, lactate metabolism were not significantly different from the control. The volummetric air flow rate had the crucial effect on the cell growth. In the large volummetric air flow rate, the cell was significantly damaged and maybe changed the requirement for nutrient. At the condition of meeting the cell oxygen demand, one-order cell death rate model was suggested and the factors concerning the death rate were discussed according to this model. 3. The pro-UK yield 1065IU/ml and 1120IU/10~6 cells at cell basis in sf9 insect cell-baculovirus expression system were achieved. The pro-UK expression level increased significantly when the cell was at good state because the yield at cell basis increased significantly and there were more cells to be infected at the same nutrient depth. Therefore, the good cell state was very important to achieve the high expression level. 4. In the hydrostatic pressure oscillation of the amplitude 0.007-0.45kgf/cm~2 and cycle 4min(2min high pressure and 2min low pressure), sf9 insect cell growth and the metabolism of glucose and lactate were insignificantly different from the control. So did the Pro-UK expression.
语种	中文
公开日期	2014-06-25
页码	90
内容类型	学位论文
源URL	[http://ir.ipe.ac.cn/handle/122111/8313]
专题	过程工程研究所_研究所（批量导入）
推荐引用方式 GB/T 7714	孙洪亮. sf9 昆虫细胞悬浮培养的研究[D]. 中国科学院研究生院. 1998.

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