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四脊滑螯虾造血组织细胞培养条件的优化
刘林敏,阮灵伟,黄智豪,施泓
0
(自然资源部第三海洋研究所、自然资源部海洋生物遗传资源重点实验室,福建 厦门361005)
摘要:
稳定的体外细胞培养体系,可以为基因功能和宿主病原相互作用研究提供平台。本研究通过对四脊滑螯虾(Cherax quadricarinatus)造血组织(hematopoietic tissue, HPT)细胞的体外培养条件进行优化,以期获得状态稳定、活力良好的造血组织细胞培养物。我们从四脊滑螯虾中分离提取造血组织细胞后,通过对培养基种类(L-15、Express Five、DMEM),谷氨酰胺浓度(0、1%、5%、10%),胎牛血清浓度(0、1%、5%、10%),以及体外培养温度(20、27 ℃)等培养条件进行优化。细胞形态观察和细胞活力分析结果表明,HPT细胞在添加1%青霉素链霉素、10%L-谷氨酰胺、1%胎牛血清的L-15培养基中状态最佳,在20 ℃培养温度下能存活30 d左右。这些培养条件将有利于四脊滑螯虾造血组织细胞的体外培养。
关键词:  海洋生物学  造血组织细胞  原代细胞  优化培养  四脊滑螯虾
DOI:10.3969/J.ISSN.2095-4972.20230220001
基金项目:福建省自然科学基金(2020J01105);现代农业产业技术体系专项资金(CARS-48)
Optimization culture of hematopoietic tissue cells of Cherax quadricarinatus
LIU Linmin,RUAN Lingwei,HUANG Zhihao,SHI Hong
(Key Laboratory of Marine Genetic Resources, Third Institute of Oceanography, MNR, Xiamen 361005, China)
Abstract:
A stable cell culture system in vitro can provide a platform for the study of gene function and host-pathogen interaction. In this study, the culture conditions of hematopoietic tissue (HPT) cells of crayfish (Cherax quadricarinatus) were optimized to generate stable HPT culture with high viability. After isolating and extracting HPT cells from crayfish, we compared the culture conditions, such as culture medium (L-15, Express Five and DMEM), glutamine concentration (0, 1%, 5% and 10%), fetal bovine serum concentration (0, 1%, 5% and 10%), and culture temperature (20 ℃ and 27 ℃). Results of cell morphology observation and cell viability analysis showed that the cell culture of L-15 medium was optimized by supplemented with 1% penicillin-streptomycin, 10% L-glutamine and 1% fetal bovine serum, in which the cells could survive for about 30 days at 20 ℃. The culture conditions are helpful for in vitro culture of hematopoietic tissue cells of crayfish.
Key words:  marine biology  hematopoietic tissue cell  primary cell  optimized culture  Cherax quadricarinatus

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