关键词： Vero细胞   动物细胞培养   球转球   接种  

摘要： 研究了静态球转球接种Ｖｅｒｏ 细胞培养过程中的细胞生长特性，提出了细胞球转球的机理，通过与消化接种的比较，证明在静态球转球接种的细胞培养过程中，细胞在新球与老球表面的生长和细胞的生理状态很不平衡，因而认为不适合于大规模细胞培养过程，并对消化接种在大规模操作中的可行性进行了讨论

关键词： biochemical engineering   cell death   filtration   DNA chromatography   fluorchrome  

摘要： The focus of this research was defined by a poorly characterised filtration train employed to clarify culture broth containing monoclonal antibodies secreted by GS-NSO cells: the filtration train blinded unpredictably and the ability of the positively charged filters to adsorb DNA from process material was unknown. To direct the development of an assay to quantify the ability of depth filters to adsorb DNA, the molecular weight of DNA from a large-scale, fed-batch, mammalian cell culture vessel was evaluated as process material passed through the initial stages of the purification scheme. High molecular weight DNA was substantially cleared from the broth after passage through a disc stack centrifuge and the remaining low molecular weight DNA was largely unaffected by passage through a series of depth filters and a sterilising grade membrane. Removal of high molecular weight DNA was shown to be coupled with clarification of the process stream. The DNA from cell culture supernatant showed a pattern of internucleosomal cleavage of chromatin when fractionated by electrophoresis but the presence of both necrotic and apoptotic cells throughout the fermentation meant that the origin of the fragmented DNA could not be unequivocally determined. An intercalating fluorochrome, PicoGreen, was elected for development of a suitable DNA assay because of its ability to respond to low molecular weight DNA. It was assessed for its ability to determine the concentration of DNA in clarified mammalian cell culture broths containing pertinent monoclonal antibodies. Fluorescent signal suppression was ameliorated by sample dilution or by performing the assay above the pI of secreted IgG. The source of fluorescence in clarified culture broth was validated by incubation with RNase A and DNase I. At least 89.0 % of fluorescence was attributable to nucleic acid and pre-digestion with RNase A was shown to be a requirement for successful quantification of DNA in such samples. Application of the

关键词： 细胞培养   大规模   动物细胞  

摘要： 大规模动物细胞培养在生物技术产业化进程中显示出强大的潜力。本文综述了大规模动物细胞培养过程中出现的问题及其解决办法 ,包括细胞培养环境、基因工程途径改建细胞系及过程监控等。对于这些进展的充分了解对优化细胞培养工艺、提高产品质量具有重要意义

关键词： 免疫原性   鸡胚   细胞培养   培养(生物)   哺乳动物细胞   菌苗   疫苗  

摘要： 试验用疫苗为双价流感A/B裂解病毒颗粒疫苗,由在MDCK细胞克隆的细胞系传代的病毒制备而成,含A/得克萨斯/36/91(类HINI)和B/哈尔滨/7/94病毒株。对照疫苗为在鸡胚传代的裂解病毒颗粒,其病毒株与试验疫苗相同。接种对象为18～50岁健康成人志愿者。按随机、双育法进行试验。将接种对象分为两组,分别接种细胞培养和鸡胚传代的流感疫苗,在三角肌肌注流感疫苗0.5ml。在接种前后分别检测血凝抑制抗体。在接种后观察不良反应。

关键词： 动物细胞   细胞培养   细胞凋亡  

摘要： 细胞培养过程中的细胞凋亡是细胞受环境因素的影响而发生的现象。随着对细胞凋亡的分子生物学和细胞生物学了解的深入，显示了有效地控制动物细胞培养中细胞凋亡的巨大潜力。包括采用ＤＮＡ重组技术把抗细胞凋亡的基因导入细胞和在培基中加入具有抗细胞凋亡的生存因子或化合物等手段已用于控制细胞培养过程中的细胞凋亡。这些技术将大大延长细胞达到饱和密度后的培养时间，提高细胞培养系统的生产效率。

关键词： 多孔载体   动物细胞培养   性质   制备材料   制备方法  

摘要： 多孔载体是一种新型的用于动物细胞培养的优秀的细胞支持物，其内部网状结构的小孔具有固定细胞和保护细胞免受机械损伤的功能，适合于贴壁细胞和悬浮细胞的培养，能提高培养密度，可应用于大规模培养系统。本文综述了多孔载体的物化性质、制作材料和制备方法。

关键词： mammalian cell culture   economy of scale   expression system   metabolic design  

摘要： Innovations for economic optimization of manufacturing processes of mammalian cell culture processes address new expression systems, optimized cell culture media and feeding systems, economy of scale, efficient harvest systems for viable cell separation, redesign of downstream processing and reduction of the overall number of quality control assays. A very efficient expression system in Chinese hamster ovary cells is the NEOSPLA expression system yielding 60-100 mu g monoclonal antibodies per cell and day. Efficient supplements in nutrient feeding are insulin and amino acids which contribute to a high extent to the productivity of the mammalian cell culture process. Large scale manufacturing processes lower cost of goods by reduction of turn around cost for cleaning and steaming in place, media preparation, number of batches for annual campaign, in-process and quality control. In downstream processing the number of process steps and the step yield are responsible for the economics. Process control systems in a computer assisted manufacturing plant increase success rate, reduces man power and minimizes shifts. In the innovative process also alternative technologies such as transgenic animals should be considered to improve the economy of the manufacturing processes.

关键词： 动物细胞   细胞培养   细胞自然凋亡  

摘要： 细胞培养过程中的细胞自然凋亡是细胞受环境压力的影响而发生的现象。随着细胞自然凋亡的分子生物学和生物化学研究的深入,对以动物细胞产品生产为目的的细胞培养产业将产生极有价值的影响。采用DNA重组技术把预防细胞自然凋亡的基因导入细胞和在培基中加入具有抗细胞自然凋亡的化合物等手段已用于预防或减缓细胞培养过程中的细胞自然凋亡。这些技术将大大延长细胞达到饱和密度后的培养时间,从而使细胞培养系统的生产效率得以显著提高。

关键词： 细胞培养   细胞凋亡   生物反应器   哺乳动物  

摘要： 目前对哺乳动物细胞培养仍进行着大量研究.最近在细胞生理学,尤其是影响细胞生长或活力的因素方面已取得巨大进展.细胞经分子遗传学操作后,更易于在无动物蛋白的培养液中培养.如何获得高细胞浓度和高细胞活力仍是细胞工程研究的共同课题.对流加培养(fed-batch culture)和灌注培养控制策略的需求,推动了工艺监测和控制的研究.上述进展对于确保制品质量和工艺稳定性十分有益.

关键词： 动物   细胞培养   大孔微载体