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含细胞的聚乙二醇水凝胶微阵列的冷冻保存

Cryopreservation of cell-containing poly(ethylene) glycol hydrogel microarrays.

作者信息

Itle Laura J, Pishko Michael V

机构信息

The Huck Institute for the Life Sciences and the Department of Chemical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802-4400, USA.

出版信息

Biotechnol Prog. 2005 May-Jun;21(3):1004-7. doi: 10.1021/bp049551+.

Abstract

Here we describe the fabrication and preservation of mammalian cell-containing hydrogel microarrays that have potential applications in drug screening and pathogen detection. Hydrogel microstructures containing murine fibroblasts were fabricated on silicon substrates and subjected to a "stage-down" freezing process. The percent viability of both immortal and primary embryonic murine fibroblast cells within the gels was determined at various stages in the freezing process, showing that cells entrapped in hydrogel microstructures remained viable throughout the process. When compared to immortalized adherent cultures subjected to the same freezing process, cells within hydrogel structures had higher cell viabilities at all stages during preservation. Finally, the necessity of using a cryoprotectant, dimethyl sulfoxide (DMSO), was investigated. Cells in hydrogels were cryopreserved with and without DMSO. The addition of DMSO altered cell viability after the freeze-thaw process, enhancing viability in an immortalized cell line and decreasing viability in a primary cell line.

摘要

在此,我们描述了含哺乳动物细胞的水凝胶微阵列的制备和保存方法,这些微阵列在药物筛选和病原体检测方面具有潜在应用价值。在硅基片上制备了含有小鼠成纤维细胞的水凝胶微结构,并对其进行“逐步降温”冷冻处理。在冷冻过程的不同阶段测定了凝胶内永生和原代胚胎小鼠成纤维细胞的存活百分比,结果表明,包裹在水凝胶微结构中的细胞在整个过程中均保持存活。与经过相同冷冻处理的永生贴壁培养物相比,水凝胶结构内的细胞在保存过程的所有阶段都具有更高的细胞活力。最后,研究了使用冷冻保护剂二甲基亚砜(DMSO)的必要性。水凝胶中的细胞在有或没有DMSO的情况下进行冷冻保存。添加DMSO会改变冻融过程后的细胞活力,提高永生细胞系中的活力,降低原代细胞系中的活力。

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