著者

菅野 正也 林 正和 村勢 則郎 網野 比佐子 北 潔

出版者

低温生物工学会

雑誌

低温生物工学会誌 (ISSN:13407902)

巻号頁・発行日

vol.54, no.2, pp.177-180, 2008-12-30 (Released:2017-06-19)

参考文献数

5

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For the establishment of successful cryopreservation of Caenorhabditis elegans, freezing survival of the nematode was investigated in the presence of dimethylsulfoxide. When nematodes were cooled to -20 ℃ with the rate of 0.2 ℃/min after ice seeding treatment at -3 ℃, survival rate of the nematode increased with the time of exposure to the DMSO solution before freezing. The rate was above 80% irrespective of the cooling temperature between -20 to -75 ℃. However, the nematodes hardly survived when they were quenched by liquid nitrogen after the pre-freezing at -20 ℃, The rate of freezing survival became higher with the decrease in the pre-freezing temperature before plunging into LN_2. Heating rate also affected freezing survival. Slow heating probably prevented rapid rehydration of the nematode and increased survival rate after thawing.

著者

林 正和 網野 比佐子 北 潔 村勢 則郎

出版者

低温生物工学会

雑誌

低温生物工学会誌 (ISSN:13407902)

巻号頁・発行日

vol.57, no.1, pp.87-90, 2011-04-15

参考文献数

8

被引用文献数

1

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Cryopreservation of the nematode Caenorhabditis elegans in the adult stage is of importance as the nematode is a powerful model organism. In this study, we applied the protocol previously established for cryopreservation of the L4 nematode to the adult one. Survival rate of 84% was obtained as a result. When ice nucleation active bacteria, Pseudomonas syringae, was used for the simplification of ice seeding procedure instead of the use of a pre-cooled needle, survival rate of 82% was obtained after thawing. Moreover, a simple method of cryopreservation using a deep freezer and polystyrene foam was developed for a practical purpose. According to the observation under a cryomicroscope, eggs in the nematode body were found to freeze at ca. -20℃ when cooled at the rate of 0.2℃/min after ice seeding at -3℃ although the nematode body remained unfrozen down to -100℃.