A cell line resource derived from honey bee (Apis mellifera) embryonic tissues.
A cell line resource derived from honey bee (Apis mellifera) embryonic tissues.
复制标题
DOI：
10.1371/journal.pone.0069831
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kurtti TJ
中科院分区：
文献类型：
--
作者：
Goblirsch MJ;Spivak MS;Kurtti TJ
研究蜜蜂病原体或杀虫剂和营养缺乏的影响的主要障碍是缺乏由蜜蜂细胞组成的受控体外培养系统。这些系统对于确定这些应激因素对蜜蜂发育和细胞生物学的影响是重要的。我们已经开发了一种方法，结合建立昆虫细胞培养技术，支持蜜蜂细胞在体外持续生长。我们使用胚胎发育中后期的蜜蜂卵来建立原代培养物，因为这些卵含有逐渐分裂的细胞。在改良的Leibovitz's L15培养基中起始原代培养物，并在32°C下孵育。维持来自几种原代培养物的材料的连续转移，并导致分离年轻细胞系。已经建立了主要由形成粘附单层的成纤维细胞型细胞组成的细胞系（AmE-711）。该品系中的大多数细胞是二倍体（2n = 32），并且具有意大利蜜蜂（Apis mellifera）核型，如Giemsa染色所示。该细胞系中的细胞色素编码的细胞色素c氧化酶亚基I（考克斯1）基因的部分序列与蜜蜂组织中的相同，并且是A.意利费拉。种群倍增时间约为4天。重要的是，当以1：3的比例分裂时，细胞系每10-14天连续传代培养，并在液氮中冷冻保存。我们已经开发的细胞培养系统具有潜在的应用研究，旨在蜜蜂的发展，遗传学，发病机制，转基因，毒理学。
A major hindrance to the study of honey bee pathogens or the effects of pesticides and nutritional deficiencies is the lack of controlled in vitro culture systems comprised of honey bee cells. Such systems are important to determine the impact of these stress factors on the developmental and cell biology of honey bees. We have developed a method incorporating established insect cell culture techniques that supports sustained growth of honey bee cells in vitro. We used honey bee eggs mid to late in their embryogenesis to establish primary cultures, as these eggs contain cells that are progressively dividing. Primary cultures were initiated in modified Leibovitz’s L15 medium and incubated at 32°C. Serial transfer of material from several primary cultures was maintained and has led to the isolation of young cell lines. A cell line (AmE-711) has been established that is composed mainly of fibroblast-type cells that form an adherent monolayer. Most cells in the line are diploid (2n = 32) and have the Apis mellifera karyotype as revealed by Giemsa stain. The partial sequence for the mitochondrial-encoded cytochrome c oxidase subunit I (Cox 1) gene in the cell line is identical to those from honey bee tissues and a consensus sequence for A. mellifera. The population doubling time is approximately 4 days. Importantly, the cell line is continuously subcultured every 10–14 days when split at a 1:3 ratio and is cryopreserved in liquid nitrogen. The cell culture system we have developed has potential application for studies aimed at honey bee development, genetics, pathogenesis, transgenesis, and toxicology.
登录
查看更多内容
影响因子：
3.7
作者：
HAYFLICK, L;MOORHEAD, PS
通讯作者：
MOORHEAD, PS
影响因子：
2.6
作者：
GASCUEL, J;MASSON, C;BEADLE, DJ
通讯作者：
BEADLE, DJ
影响因子：
11
作者：
Huang HL;Hsing HW;Lai TC;Chen YW;Lee TR;Chan HT;Lyu PC;Wu CL;Lu YC;Lin ST;Lin CW;Lai CH;Chang HT;Chou HC;Chan HL
通讯作者：
Chan HL
影响因子：
2.6
作者：
Chan, M. M. Y.;Choi, S. Y. C.;Foster, L. J.
通讯作者：
Foster, L. J.
DOI：
10.1007/s11626-011-9431-6
发表时间：
2011-08-01
影响因子：
2.1
作者：
Kitagishi, Yasuko;Okumura, Naoko;Matsuda, Satoru
通讯作者：
Matsuda, Satoru