CHO细胞无血清培养基的筛选与优化

发布时间：2018-06-27 08:29

本文选题：CHO细胞 + 细胞培养　；参考：《吉林大学》2017年硕士论文

【摘要】：目的:动物细胞培养技术是当今生物工程领域中的前沿技术之一,无论是在研究还是在生产实践方面都对当代生物技术有着重要的影响。虽然目前市售的商业化培养基已具有非常好的广谱性,但是却缺少针对性。针对某一细胞品种设计对应的培养基成分无疑是需要耗费大量的金钱和时间,利用多种市售的培养基进行不同比例的混合,可以在较短时间内使细胞培养得到针对性优化,从而提高产物的产量。方法:本文利用了市售的几种常用培养基,对CHO细胞进行培养,采用先筛选后优化组合的方式进行针对文中的CHO细胞培养进行优化。筛选和优化又分为两部分进行:一是基础培养基的筛选和优化;二是补料培养基的筛选和优化。在培养过程中检测细胞的活率、密度以及产物的表达量作为培养基的评价指标,用以考察培养基的优劣。结果:通过对基础培养基和补料培养基的筛选、优化,使得最终的利用混合培养基培养的细胞不论是在细胞密度、活率还是产物蛋白的表达量方面都优于单一培养基培养的细胞。通过优化使得基础培养基筛在最细胞高密度方面:混合培养基的平均最高密度高出单一培养基的平均最高密度9.8%;在最后细胞活率方面,混合培养基的最终平均活率高出单一培养基的最终平均活率11.8%;在最终蛋白产量方面,混合培养基的最终产量高出单一培养基的最终产量17.4%。通过优化补料培养基在最细胞高密度方面:混合培养基的平均最高密度高出单一培养基的平均最高密度11.0%;在最后细胞活率方面,混合培养基的最终平均活率高出单一培养基的最终平均活率26.6%;在最终蛋白产量方面,混合培养基的最终产量高出单一培养基的最终产量20.6%。结论通过初步筛选和优化组合的方式进行多种培养基的成分配比可以在短时间内提高细胞密度、提升细胞活率的维持时间从而提高目的蛋白的产量。
[Abstract]:Objective: animal cell culture technology is one of the frontier technologies in the field of bioengineering. It has important influence on modern biotechnology in both research and production practice. Although the commercial media on the market has a very good broad spectrum, but the lack of pertinence. It is necessary to design the corresponding culture medium components for a certain cell variety. It takes a lot of money and time to mix the different proportion of the medium sold on the market, which can optimize the cell culture in a short period of time. Thus, the output of the product is increased. Methods: Cho cells were cultured on several commonly used culture media in this paper. Cho cell culture was optimized by first screening and then optimizing combination. Screening and optimization are divided into two parts: one is the screening and optimization of the basic medium, the other is the screening and optimization of the supplement medium. In the process of culture, the cell viability, density and the expression of the product were measured as the evaluation indexes of the culture medium to evaluate the quality of the medium. Results: by screening and optimizing the basic medium and the supplement medium, the cells cultured in the mixed medium could be cultured in the cell density. Both the viability and the expression of protein were superior to those of the cells cultured on a single medium. By optimizing the basic medium sieve, the cell density is the highest: the average maximum density of the mixed medium is higher than the average maximum density of the single medium, and the final cell viability is 9.80% higher than the average maximum density of the single medium. The final average survival rate of the mixed medium was higher than that of the single medium, and the final yield of the mixed medium was 17.4% higher than that of the single medium, and the final protein yield of the mixed medium was 17.4% higher than that of the single medium. The maximum cell density of the mixed medium was higher than that of the single medium, and the final cell viability was higher than that of the single medium. The final average survival rate of the mixed medium was higher than that of the single medium, and the final yield of the mixed medium was 20.6 higher than that of the single medium. Conclusion by preliminary screening and optimizing combination, the composition ratio of various media can increase cell density and maintain time of cell viability in a short time, so as to increase the yield of target protein.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q813.11

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