Watanabe Kazuyo, Imanishi Shigeo, Akiduki Gaku, Cornette Richard, Okuda Takashi
Institute of Agrobiological Sciences, NARO, 1-2 Ohwashi, Tsukuba, Ibaraki, 305-8634, Japan.
Institute of Agrobiological Sciences, NARO, 1-2 Ohwashi, Tsukuba, Ibaraki, 305-8634, Japan.
Cryobiology. 2016 Aug;73(1):93-8. doi: 10.1016/j.cryobiol.2016.05.006. Epub 2016 May 17.
Pv11, a cell line derived from the anhydrobiotic insect, Polypedilum vanderplanki, was preserved in a dry form (only 6% residual moisture) at room temperature for up to 251 days and restarted proliferating after rehydration. A previous study already reported survival of Pv11 cells after desiccation, but without subsequent proliferation. Here, the protocol was improved to increase survival and achieve proliferation of Pv11 cells after dry storage. The method basically included preincubation, desiccation and rehydration processes and each step was investigated. So far, preincubation in a 600 mM trehalose solution for 48 h before dehydration was the most favourable preconditioning to achieve successful dry preservation of Pv11 cells, allowing about 16% of survival after rehydration and subsequent cell proliferation. Although the simple air-dry method established for Pv11 cells here was not applicable for successful dry-preservation of other insect cell lines, Pv11 is the first dry-preservable animal cell line and will surely contribute not only to basic but also applied sciences.
Pv11是一种源自脱水生物昆虫范氏摇蚊(Polypedilum vanderplanki)的细胞系,以干燥形式(仅6%的残余水分)在室温下保存长达251天,复水后重新开始增殖。先前的一项研究已经报道了Pv11细胞在干燥后的存活情况,但没有后续的增殖。在此,对方案进行了改进,以提高干燥保存后Pv11细胞的存活率并实现其增殖。该方法基本包括预孵育、干燥和复水过程,并且对每个步骤都进行了研究。到目前为止,在脱水前于600 mM海藻糖溶液中预孵育48小时是实现Pv11细胞成功干燥保存的最有利预处理条件,复水后约有16%的存活率以及随后的细胞增殖。尽管此处为Pv11细胞建立的简单风干方法不适用于其他昆虫细胞系的成功干燥保存,但Pv11是第一个可干燥保存的动物细胞系,无疑将不仅对基础科学而且对应用科学做出贡献。