Injury of larger biological tissue by extracellular freezing

Masanobu Ujihira*, Ryou Yamaguchi, Naoki Aizawa, Kazuo Tanishita

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


The purpose of this study is to reduce injury of larger cells and tissue (≥ 1 mm) due to extracellular freezing to achieve successful cryopreservation. In the temperature range of 0 to -40°C, the morphology of the fertilized killifish egg was observed under a microscope with a cooling rate from 0.1 to 10°C/min. In glycerol-water, DMSO-water (cryoprotectant solution) and distilled water, the damage rate to the egg by maintaining extracellular freezing of short duration of various temperatures was evaluated by hatching rate. As a result, when the egg shell defectively buckled due to dehydration of perivitelline, the hatching rate was more than 80% in glycerol-water solution. Hatching rate was maximum at the glycerol concentration of 7.5%. Nearly identical results were obtained in DMSO-water solution (maximum at 15%). In distilled water, hatching rate was very low. Transformation of egg shell and injury of the egg are not correlated. Optimum concentration of cryoprotectant minimizes injury of larger cells and tissue due to extracellular freezing.

Original languageEnglish
Pages (from-to)3066-3074
Number of pages9
JournalNippon Kikai Gakkai Ronbunshu, B Hen/Transactions of the Japan Society of Mechanical Engineers, Part B
Issue number588
Publication statusPublished - 1995 Aug

ASJC Scopus subject areas

  • Mechanical Engineering


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