Orbital shaker technology for the cultivation of mammalian cells in suspension
Orbital shaker technology for the cultivation of mammalian cells in suspension
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DOI：
10.1002/bit.20358
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发表时间：
2005-02-20
影响因子：
3.8
通讯作者：
Wurm, FM
中科院分区：
文献类型：
--
作者：
Muller, N;Girard, P;Wurm, FM
为了大规模应用于生物技术，哺乳动物细胞的悬浮培养是必不可少的先决条件。通常，悬浮培养是在玻璃旋转烧瓶中生长，填充到标称体积的50%以下。我们提出了一种基于轨道振动器技术的哺乳动物细胞悬浮培养系统。我们发现“方形”瓶（方瓶）提供了一种廉价但有效的方法来培养HEK-293 EBNA和CHO-DG44细胞到高密度。1-L方形瓶中的培养物的性能超过了旋转瓶中的培养物，HEK-293 EBNA细胞的密度高达7 × 10(6)个细胞/mL， CHO-DG44细胞的密度高达5 × 10(6)个细胞/mL，而在旋转瓶中培养的相同细胞的密度为（2.5 - 4）× 106个细胞/mL。对于1 l的方形瓶，当填充体积为标称体积的30-40%，搅拌速度为130 rpm，旋转直径为2.5 cm时，观察到最佳的细胞生长和活力。磷酸钙- DNA共沉淀法传递基因后，方形瓶中生长的HEK-293 EBNA细胞的瞬时报告基因表达与旋转瓶中生长的相同或稍好。降低成本、简化操作、更好的细胞生长性能和活力使搅拌方形瓶成为一种新的、非常有前途的哺乳动物细胞悬浮培养工具。(C) 2004 Wiley期刊有限公司
For large-scale applications in biotechnology, cultivation of mammalian cells in suspension is an essential prerequisite. Typically, suspension cultures are grown in glass spinner flasks filled to less than 50% of the nominal volume. We propose a superior system for suspension cultures of mammalian cells based on orbital shaker technology. We found that "square-shaped" bottles (square bottles) provide an inexpensive but efficient means to grow HEK-293 EBNA and CHO-DG44 cells to high density. Cultures in agitated 1-L square bottles exceeded the performance of cultures in spinner flasks, reaching densities up to 7 x 10(6) cells/mL for HEK-293 EBNA cells and 5 x 10(6) cells/mL for CHO-DG44 cells in comparison to (2.5 - 4) x 106 cells/mL for cultures of the same cells grown in spinner flasks. For 1-L square bottles, optimal cell growth and viability were observed with a filling volume of 30-40% of the nominal volume and an agitation speed of 130 rpm at a rotational diameter of 2.5 cm. Transient reporter gene expression following gene delivery by calcium phosphate - DNA co-precipitation was the same or slightly better for HEK-293 EBNA cells grown in square bottles as compared to spinner flasks. Reductions in cost, simplified handling, and better performance in cell growth and viability make the agitated square bottle a new and very promising tool for the cultivation of mammalian cells in suspension. (C) 2004 Wiley Periodicals, Inc.