Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification

Micro-droplet spray vitrification system produces small droplets and can be vitrified at a lower concentration of cryoprotectant (CPA). In this study, HepG2 cells were vitrified using micro-droplet vitrification system; moreover, damages caused by CPA loading, spraying, receiving, and vitrification/...

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Main Authors: Peng Tian, Zhang Xiaomin, Zhou Xinli
Format: Article
Language:zho
Published: Journal of Refrigeration Magazines Agency Co., Ltd. 2020-01-01
Series:Zhileng xuebao
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Online Access:http://www.zhilengxuebao.com/thesisDetails#10.3969/j.issn.0253-4339.2020.02.152
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author Peng Tian
Zhang Xiaomin
Zhou Xinli
author_facet Peng Tian
Zhang Xiaomin
Zhou Xinli
author_sort Peng Tian
collection DOAJ
description Micro-droplet spray vitrification system produces small droplets and can be vitrified at a lower concentration of cryoprotectant (CPA). In this study, HepG2 cells were vitrified using micro-droplet vitrification system; moreover, damages caused by CPA loading, spraying, receiving, and vitrification/rewarming processes were evaluated. In addition, CPA formulations were optimized by reducing the Me2SO concentration and adding an appropriate amount of trehalose. The results showed that each step in micro-droplet spray vitrification may cause damage to the cells. CPA loading, spraying, vitrification and rewarming processes caused severe damages, but mild damages were caused by the sheet receiving process. Cell activities after cryopreservation are significantly reduced with a decrease in the Me2SO concentration. When the CPA formulations are constant, cell activities in the vitrification group become better than those in the slow freezing group. Moderate amounts of trehalose can enhance the effect of cryopreservation, while excessive trehalose can be counteractive. When cells were preserved with 5% Me2SO + 0.3M trehalose as a cryoprotectant, the cell survival rate reached (92.42±0.95)% and the 24 h adherence rate reached (95.64±1.03)%; therefore, the droplet spray vitrification effect was found to be the best.
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issn 0253-4339
language zho
publishDate 2020-01-01
publisher Journal of Refrigeration Magazines Agency Co., Ltd.
record_format Article
series Zhileng xuebao
spelling doaj-art-4fd214e72e704ab3adb2c41b681c433b2025-08-20T03:15:53ZzhoJournal of Refrigeration Magazines Agency Co., Ltd.Zhileng xuebao0253-43392020-01-014166509447Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray VitrificationPeng TianZhang XiaominZhou XinliMicro-droplet spray vitrification system produces small droplets and can be vitrified at a lower concentration of cryoprotectant (CPA). In this study, HepG2 cells were vitrified using micro-droplet vitrification system; moreover, damages caused by CPA loading, spraying, receiving, and vitrification/rewarming processes were evaluated. In addition, CPA formulations were optimized by reducing the Me2SO concentration and adding an appropriate amount of trehalose. The results showed that each step in micro-droplet spray vitrification may cause damage to the cells. CPA loading, spraying, vitrification and rewarming processes caused severe damages, but mild damages were caused by the sheet receiving process. Cell activities after cryopreservation are significantly reduced with a decrease in the Me2SO concentration. When the CPA formulations are constant, cell activities in the vitrification group become better than those in the slow freezing group. Moderate amounts of trehalose can enhance the effect of cryopreservation, while excessive trehalose can be counteractive. When cells were preserved with 5% Me2SO + 0.3M trehalose as a cryoprotectant, the cell survival rate reached (92.42±0.95)% and the 24 h adherence rate reached (95.64±1.03)%; therefore, the droplet spray vitrification effect was found to be the best.http://www.zhilengxuebao.com/thesisDetails#10.3969/j.issn.0253-4339.2020.02.152micro-droplet sprayvitrificationprocess damagescryoprotecant optimization
spellingShingle Peng Tian
Zhang Xiaomin
Zhou Xinli
Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
Zhileng xuebao
micro-droplet spray
vitrification
process damages
cryoprotecant optimization
title Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
title_full Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
title_fullStr Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
title_full_unstemmed Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
title_short Process Damage Analysis and Cryoprotectant Optimization for Micro-droplet Spray Vitrification
title_sort process damage analysis and cryoprotectant optimization for micro droplet spray vitrification
topic micro-droplet spray
vitrification
process damages
cryoprotecant optimization
url http://www.zhilengxuebao.com/thesisDetails#10.3969/j.issn.0253-4339.2020.02.152
work_keys_str_mv AT pengtian processdamageanalysisandcryoprotectantoptimizationformicrodropletsprayvitrification
AT zhangxiaomin processdamageanalysisandcryoprotectantoptimizationformicrodropletsprayvitrification
AT zhouxinli processdamageanalysisandcryoprotectantoptimizationformicrodropletsprayvitrification