Cultivating cells in shake flasks is a routine operation that is largely unchanged since its inception. A glass or plastic Erlenmeyer vessel with the primary gas exchange taking place across various porous plugs is used with media volumes typically ranging from 100 mL to 2 L. Oxygen limitation and carbon dioxide accumulation in the vessel is a major concern for studies involving shake flask cultures. In this study, we enhance mass transfer in a conventional shake flask by replacing the body wall with a permeable membrane. Naturally occurring concentration gradient across the permeable membrane walls facilitates the movement of oxygen and carbon dioxide between the flask and the external environment. The modified flask called the breathable flask, has shown a 40% improvement in mass transfer coefficient (kLa) determined using the static diffusion method. The prokaryotic cell culture studies performed with Escherichia coli showed an improvement of 28%–66% in biomass and 41%–56% in recombinant product yield. The eukaryotic cell culture study performed with Pichia pastoris expressing proinsulin exhibited a 40% improvement in biomass and 115% improvement in protein yield. The study demonstrates a novel approach to addressing the mass transfer limitations in conventional shake flask cultures. The proposed flask amplifies its value by providing a membrane‐diffusion‐based sensing platform for the integration of low‐cost, noninvasive sensing capabilities for real‐time monitoring of critical cell culture parameters like dissolved oxygen and dissolved carbon dioxide.

中文翻译：

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重塑摇瓶发酵：透气瓶

在摇瓶中培养细胞是一种常规操作，自诞生以来基本没有变化。使用玻璃或塑料锥形容器，主要气体交换发生在各种多孔塞上，培养基体积通常为 100 mL 至 2 L。容器中的氧气限制和二氧化碳积累是涉及摇瓶培养的研究的主要问题。在这项研究中，我们通过用渗透膜取代体壁来增强传统摇瓶中的传质。渗透膜壁上自然发生的浓度梯度有利于氧气和二氧化碳在烧瓶和外部环境之间的移动。改良后的烧瓶称为透气烧瓶，其传质系数提高了 40%（kLA）使用静态扩散法测定。原核细胞培养研究大肠杆菌生物量提高了 28%–66%，重组产物产量提高了 41%–56%。真核细胞培养研究采用毕赤酵母表达胰岛素原的生物量提高了 40%，蛋白质产量提高了 115%。该研究展示了一种解决传统摇瓶培养中传质限制的新方法。所提出的烧瓶通过提供基于膜扩散的传感平台来集成低成本、非侵入性传感功能，从而实时监测溶解氧和溶解二氧化碳等关键细胞培养参数，从而扩大了其价值。