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A Simple Mechanistic Way to Increase the Survival of Mammalian Cells During Processing for Dry Storage

机译：一种简单的机械方法，可以增加干细胞加工过程中哺乳动物细胞的存活率

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Recently, there has been considerable interest in developing processing methods that enable storage of cells in a dry state. Most of these studies describe cell viability following processing as a function of the final moisture content reached or the duration of drying. Recently, a cumulative osmotic stress model has been proposed, which takes both final moisture content and duration of drying into consideration in an effort to account for the effects of cumulative processing stresses. The present study demonstrates the applicability of this approach and elucidates a simple mechanistic technique to reduce cumulative osmotic stress during processing. Mouse macrophage cells ( J774) were exposed to increasing concentrations of trehalose-containing 0.33phosphatebuffered saline (PBS) by step-changing the extracellular solution in 2 increments of 0.7 Osm, using only trehalose as the additive solute. Three minutes was provided for equilibration prior to drying in a traditional lowhumidity chamber. The data were compared with that of cells dried directly in isotonic 0.2M trehalose in 0.33 PBS. Following dehydration, cells were rehydrated and viability was assessed 45 min postrehydration using a combination of trypan blue staining for membrane integrity of detached cells and calcein AM–propidium iodide fluorescence assay for live–dead staining of the attached cells. Cells that were preprocessed to higher trehalose concentrations in step changes prior to drying had higher viability scores at comparable final moisture levels when compared with cells dried in iso-osmotic solution, up to a limit of *8 Osm, at which point cells processed by both methods approached zero viability.
机译：最近，人们对开发能够以干燥状态储存细胞的加工方法引起了极大的兴趣。这些研究大多数将处理后的细胞活力描述为达到的最终水分含量或干燥时间的函数。最近，已经提出了一种累积渗透应力模型，该模型同时考虑了最终的水分含量和干燥时间，以考虑累积加工应力的影响。本研究证明了这种方法的适用性，并阐明了一种简单的机械技术来减少加工过程中的累积渗透压。小鼠巨噬细胞（J774）仅以海藻糖为添加溶质，以2个增量（0.7渗透压）逐步改变细胞外溶液，使之暴露于浓度更高的含海藻糖的0.33磷酸盐缓冲盐水（PBS）中。在传统的低湿度室内干燥之前，需要提供三分钟的平衡时间。将数据与在等渗0.2M海藻糖的0.33 PBS中直接干燥的细胞进行比较。脱水后，将细胞再水化，并在脱水后45分钟使用台盼蓝染色（用于分离细胞的膜完整性）和钙黄绿素AM–碘化丙啶荧光测定结合细胞的活死染色的方法评估生存力。与在等渗溶液中干燥的细胞相比，在干燥前逐步改变为高阶海藻糖浓度的细胞在可比的最终水分水平下具有较高的生存力得分，最高可达* 8 Osm，此时两个细胞都处理方法接近零生存力。

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