1 Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts.
Am J Respir Cell Mol Biol. 2014 May;50(5):876-81. doi: 10.1165/rcmb.2013-0166MA.
An emerging tool in airway biology is the precision-cut lung slice (PCLS). Adoption of the PCLS as a model for assessing airway reactivity has been hampered by the limited time window within which tissues remain viable. Here we demonstrate that the PCLS can be frozen, stored long-term, and then thawed for later experimental use. Compared with the never-frozen murine PCLS, the frozen-thawed PCLS shows metabolic activity that is decreased to an extent comparable to that observed in other cryopreserved tissues but shows no differences in cell viability or in airway caliber responses to the contractile agonist methacholine or the relaxing agonist chloroquine. These results indicate that freezing and long-term storage is a feasible solution to the problem of limited viability of the PCLS in culture.
在气道生物学中,一种新兴的工具是精密切割肺切片(PCLS)。由于组织在保持活力的时间窗口有限,因此采用 PCLS 作为评估气道反应性的模型受到了阻碍。在这里,我们证明了 PCLS 可以冷冻、长期储存,然后解冻用于以后的实验使用。与从未冷冻的鼠 PCLS 相比,冷冻解冻的 PCLS 显示出代谢活性降低,程度与其他冷冻保存组织相似,但在细胞活力或气道对收缩激动剂乙酰甲胆碱或松弛激动剂氯喹的反应方面没有差异。这些结果表明,冷冻和长期储存是解决 PCLS 在培养中活力有限的问题的可行方法。