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用于细胞培养的非灌注附着系统。

Nonperfused attachment systems for cell cultivation.

作者信息

Elliott A Y

机构信息

Merck Pharmaceutical Manufacturing Division of Merck & Co., Inc., West Point, Pennsylvania.

出版信息

Bioprocess Technol. 1990;10:207-16.

PMID:1367060
Abstract

The pressing need for large-scale culture methods has prompted efforts to develop vessels which will accommodate the growth of large numbers of anchorage-dependent cells. The large-scale "cell factories" have been used to grow viruses for vaccine production, nucleic acid studies, and various cancer research projects. Two particular types of culture vessels used for large-scale production of anchorage-dependent cells were discussed and examples of their use in vaccine production given. Development of these large-scale culture systems has enabled the pharmaceutical companies to (1) meet the ever-increasing demands worldwide for vaccine, (2) employ a production process that produces a cost-efficient vaccine product in cell culture, and (3) produce large volumes of bulk vaccine at a single campaign, thus allowing multiple usage of a single production unit over the course of a year. The type of large-scale culture vessel used depends on the purpose of the culture and the type of cells that are to be grown in the vessel. The unit process vessel, with its multidisks, provides superior surface area, but because of the stainless steel housing, the cells cannot be monitored microscopically. Additionally, the unit process vessel can be equipped with a jacket for precise temperature regulation and thus eliminate the need for incubators. The roller bottles, on the other hand, allow microscopic monitoring of the growing cells but are limited in surface area available for growth, and the volume of harvest fluid obtainable from a bottle is very limited. It should be noted that handling of roller bottles is labor-intensive and requires many more manipulations and hence more personnel than employing unit process-type vessels for vaccine-manufacturing operations. The use of mass cultivation techniques in cancer research has also been of immense benefit. The production of large numbers of neoplastic cells has provided researchers with the raw materials to perform molecular biology experiments such as DNA sequencing, which required milligram quantities of cells before sufficient quantities of DNA could be isolated to allow sequencing studies. Additionally, the technique of mass cultivation also provided sufficient quantities of cells for inoculation into either nude mice or immunosuppressed hamsters in an attempt to determine the tumor induction capacity of the cultured cells. Additional characterization studies which require large numbers of cell, enzyme, and isoenzyme profiles can also be performed on cells grown in roller bottle cultures. There are numerous factors which limit the productivity of mass cultivation systems and the scientist should be aware of their existence.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

对大规模培养方法的迫切需求促使人们努力开发能够容纳大量贴壁依赖性细胞生长的容器。大规模的“细胞工厂”已被用于培养病毒以生产疫苗、进行核酸研究以及开展各种癌症研究项目。文中讨论了两种用于大规模生产贴壁依赖性细胞的特定类型培养容器,并给出了它们在疫苗生产中的使用示例。这些大规模培养系统的发展使制药公司能够:(1)满足全球对疫苗不断增长的需求;(2)采用在细胞培养中生产具有成本效益的疫苗产品的生产工艺;(3)在一次生产活动中生产大量的批量疫苗,从而使单个生产单元在一年中能够多次使用。所使用的大规模培养容器类型取决于培养目的以及要在容器中生长的细胞类型。带有多个盘片的单元处理容器提供了更大的表面积,但由于其不锈钢外壳,无法在显微镜下监测细胞。此外,单元处理容器可配备夹套以进行精确的温度调节,从而无需使用培养箱。另一方面,滚瓶允许对生长中的细胞进行显微镜监测,但可供生长的表面积有限,并且从一个瓶子中可获得的收获液体积非常有限。应当注意的是,处理滚瓶劳动强度大,与使用单元处理型容器进行疫苗生产操作相比,需要更多的操作步骤,因此需要更多的人员。大规模培养技术在癌症研究中的应用也带来了巨大益处。大量肿瘤细胞的产生为研究人员提供了进行分子生物学实验(如DNA测序)的原材料,在能够分离出足够数量的DNA以进行测序研究之前,DNA测序需要毫克级数量的细胞。此外,大规模培养技术还提供了足够数量的细胞用于接种到裸鼠或免疫抑制的仓鼠体内,以试图确定培养细胞的致瘤能力。对于在滚瓶培养中生长的细胞,还可以进行需要大量细胞、酶和同工酶谱的其他表征研究。有许多因素限制了大规模培养系统的生产力,科学家应该意识到它们的存在。(摘要截选至400字)

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