Two skin cell lines from wild-type and albino Japanese flounder (Paralichthys olivaceus): establishment, characterization, virus susceptibility, efficient transfection, and application to albinism study
Two skin cell lines from wild-type and albino Japanese flounder (Paralichthys olivaceus): establishment, characterization, virus susceptibility, efficient transfection, and application to albinism study
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来自野生型和白化牙鲆（Paralichthys olivaceus）的两种皮肤细胞系：建立、表征、病毒敏感性、有效转染以及在白化病研究中的应用
DOI：
10.1007/s10695-017-0386-8
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发表时间：
2017-07
影响因子：
2.9
通讯作者：
Wang Na
中科院分区：
文献类型：
--
作者：
Wang Ruoqing;Zhang Nianwei;Wang Renkai;Wang Shengpeng;Wang Na
为了为研究鱼类病理学和皮肤色素沉着提供适用的细胞平台，建立了两个源自野生型和白化日本牙鲆皮肤组织的细胞系，分别命名为JFSK_wt和JFSK_alb。这两种细胞系在大约 300 天内培养了 45 代。 JFSK_wt 和 JFSK_alb 细胞维持在补充有抗生素、胎牛血清 (FBS)、2-巯基乙醇 (2-Me)、N-2-羟乙基哌嗪-N'-2-乙磺酸 (HEPES) 和碱性成纤维细胞生长因子 (bFGF) 的 Dulbecco 改良 Eagle 培养基和 Ham F-12 营养混合物 (DMEM/F12) 中。 JFSK_wt 和 JFSK_alb 细胞的最适生长温度为 24 °C，显微镜下观察，这两种细胞系均由成纤维细胞样细胞组成。染色体分析显示，JFSK_wt 和 JFSK_alb 细胞具有相同的二倍体核型，2n = 48t。病毒接种测定的结果显示，两种细胞系对神经坏死病毒（NNV）具有相似的病毒易感性模式。在用 pEGFP-N3 报告质粒和 Cy3-siRNA 转染的 JFSK_wt 和 JFSK_alb 细胞中观察到高转染效率。真皮标记物Dermo-1的检测表明这两种细胞均来源于真皮。最后，与 JFSK_wt 细胞相比，参与黑色素生成途径的三个基因，包括 5 型腺苷酸环化酶 (adcy5)、小眼相关转录因子 (mitf) 和内皮素 B 受体 (ednrb)，在 JFSK_alb 细胞中下调。因此，从野生型和白化牙鲆皮肤组织中采集的这两种细胞系不仅有助于鱼类病原体研究，而且有利于白化病相关基因功能研究。
In order to provide an applicable cell platform to study fish pathology and skin pigmentation, two cell lines derived from skin tissues of wild-type and albino Japanese flounder were established and named JFSK_wt and JFSK_alb, respectively. These two cell lines were cultured for 45 passages within approximately 300 days. JFSK_wt and JFSK_alb cells were maintained in Dulbecco’s Modified Eagle’s Medium and Ham’s F-12 Nutrient Mixture (DMEM/F12) supplemented with antibiotics, fetal bovine serum (FBS), 2-mercaptoethanol (2-Me),N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), and basic fibroblast growth factor (bFGF). The optimal growth temperature for JFSK_wt and JFSK_alb cells was 24 °C, and microscopically, the two cell lines were composed of fibroblast-like cells. Chromosomal analysis revealed that JFSK_wt and JFSK_alb cells had an identical diploid karyotype with 2n = 48t. Results of viral inoculation assays revealed that both cell lines shared similar patterns of viral susceptibility to nervous necrosis virus (NNV). High transfection efficiency was observed in JFSK_wt and JFSK_alb cells transfected with a pEGFP-N3 reporter plasmid and Cy3-siRNA. The detection of dermal marker Dermo-1 showed that these two cells were both derived from the dermis. Finally, three genes involved in the melanogenesis pathway, including adenylate cyclase type 5 (adcy5), microphthalmia-associated transcription factor (mitf), and endothelin B receptor (ednrb), were downregulated in JFSK_alb versus JFSK_wt cells. Thus, the two cell lines, sampled from skin tissue of wild-type and albino Japanese flounder will be not only helpful for fish pathogen research but also beneficial for albinism-related gene function studies.
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影响因子：
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作者：
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通讯作者：
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影响因子：
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