茶悬浮培养细胞玻璃化超低温保存研究

刘亚军; 高丽萍; 夏涛; 高可君

doi:10.13305/j.cnki.jts.2009.2.006

茶叶科学 >

2009 , Vol. 29 >Issue 2: 120 - 126

DOI: https://doi.org/10.13305/j.cnki.jts.2009.2.006

茶悬浮培养细胞玻璃化超低温保存研究

刘亚军 ,
高丽萍 ,
夏涛 ,
高可君

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1. 安徽农业大学茶叶生物化学与生物技术教育部、农业部重点实验室,安徽合肥 230036;
2. 安徽农业大学生命科学学院,安徽合肥 230036

刘亚军(1981— ),男,安徽合肥人,博士研究生,从事茶叶生化及天然产物研究。

收稿日期: 2008-07-18

修回日期: 2008-11-17

网络出版日期: 2019-09-06

基金资助

国家重点基础研究发展计划“973”前期项目（2007CB116211),国家自然科学基金项目（30771755）

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Study on Cryopreservation of Suspension Culture Cells by Vitrification in Camellia Sinensis

LIU Ya-jun ,
GAO Li-ping ,
XIA Tao ,
GAO Ke-jun

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1. Key Lab of Tea Biochemistry & Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China;;
2. School of Biology Science, Anhui Agricultural University, Hefei 230036, China

Received date: 2008-07-18

Revised date: 2008-11-17

Online published: 2019-09-06

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摘要

为保存具有稳定代谢能力的优良茶细胞系,本文对茶叶悬浮培养细胞玻璃化法超低温保存进行了初步研究。结果表明：预培养以4 d最佳,60% PVS₂冰浴装载以20 min最好,100% PVS₂冰浴脱水处理60 min最优,40℃复温解冻细胞存活率最高,采用这些参数进行完整试验的细胞存活率达到76%,初步建立了茶悬浮培养细胞玻璃化超低温保存的冷冻程序。

关键词： 茶; 悬浮培养细胞; 玻璃化法; 超低温保存

本文引用格式

刘亚军 , 高丽萍 , 夏涛 , 高可君 . 茶悬浮培养细胞玻璃化超低温保存研究[J]. 茶叶科学, 2009 , 29(2) : 120 -126 . DOI: 10.13305/j.cnki.jts.2009.2.006

Abstract

To preserve eminent cell lines with stable metabolism, the cryopreservation of suspension culture cells by vitrification in Camellia sinensis has been preliminarily investigated in this article. It showed that the preculture for four days was best, loading with 60% PVS₂ in freezing bath for 20 minutes was optimal. Treating with 100% PVS₂ dehydration in freezing bath for 60 minutes was suggested. The cryopreservation of suspension culture cells warmed in 40℃water bath had the highest survival rate. The cells’ survival rate was as high as 76% when these parameters were adopted through the whole trial. The procedure of cryopreservation of suspension culture cells by vitrification in Camellia sinensis was preliminarily established.

Key words： tea(Camellia sinensis); suspension culture cell; vitrification; cryoproservation

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