Shijun Zhu, Kamran Jamil, Xiaocui Ma, J. Crowe, A. E. Oliver
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引用次数: 3
摘要
CANARY细胞是经过基因工程改造的小鼠B细胞,可作为各种感染性病原体的快速检测系统。我们正在尝试生产一种稳定的脱水细胞产品,使该系统实用。这些细胞在干燥过程中受损的主要原因是细胞凋亡。海藻糖具有保护哺乳动物细胞在干燥过程中的作用,研究了海藻糖对细胞凋亡的影响。B细胞装载海藻糖并真空干燥。海藻糖在干燥和复水化过程中减少凋亡,与泛半胱天冬酶抑制剂opho -109联合使用时,凋亡细胞损失程度进一步降低。在这种情况下,补液后的存活率达到70%-80%。令人惊讶的是,单独使用海藻糖比单独使用opho -109更好地阻止凋亡细胞死亡(在干燥至0.3 g H2O/g干重后,海藻糖或opho -109处理的样品中,凋亡细胞总数分别为45%和70%)。然而,当细胞被加载和加载时,达到了最佳的生存能力。
Protection of CANARY Cells After Drying and Rehydration Correlates with Decrease in Apoptotic Cell Death
CANARY cells are genetically engineered murine B cells that serve as a rapid detection system for various infectious pathogens. We are attempting to produce a stable dehydrated cellular product to make this system practical. A major source of damage to these cells during drying was identified as apoptosis. Trehalose, which protects mammalian cells during drying, was investigated with regard to its effect on apoptosis. B cells were loaded with trehalose and vacuum-dried. Trehalose reduced apoptosis during drying and rehydration, and when used in combination with a pan-caspase inhibitor OPH-109, the degree of apoptotic cell loss was further diminished. In this case, viability following rehydration reached 70%–80%. Surprisingly, trehalose alone blocked apoptotic cell death better than OPH-109 alone (45% versus 70% total apoptotic cells for trehalose- or OPH-109–treated samples, respectively, after drying to 0.3 g H2O/g dry weight). Nevertheless, optimal viability was achieved when the cells were loaded and d...