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美国鳗鱼脑内皮样细胞系对不同暴露培养基中硒缺乏以及亚硒酸盐、硒酸盐和硒代蛋氨酸添加的反应。

Responses of an American eel brain endothelial-like cell line to selenium deprivation and to selenite, selenate, and selenomethionine additions in different exposure media.

作者信息

Bloch Sophia R, Kim John J, Pham Phuc H, Hodson Peter V, Lee Lucy E J, Bols Niels C

机构信息

Department of Biology, University of Waterloo, Waterloo, Waterloo, ON, N2L 3G1, Canada.

Department of Biology and School of Environmental Studies, Queen's University, Kingston, ON, K7L 3N6, Canada.

出版信息

In Vitro Cell Dev Biol Anim. 2017 Dec;53(10):940-953. doi: 10.1007/s11626-017-0196-4. Epub 2017 Sep 22.

Abstract

The effect of selenium deprivation and addition on the American eel brain endothelial cell line (eelB) was studied in three exposure media: complete growth medium (L15/FBS), serum-free medium (L15), and minimal medium (L15/ex). L15/ex contains only galactose and pyruvate and allowed the deprivation of selenium on cells to be studied. In L15/ex, without any obvious source of selenium, eelB cells survived for at least 7 d, formed capillary-like structures (CLS) on Matrigel, and migrated to heal wounds. Three selenium compounds were added to cultures: selenite, selenate, and selenomethionine (SeMet). Adding selenite or selenate to eelB cell cultures for 24 h caused dose-dependent declines in cell viability, regardless of the exposure media. Although varying with exposure media and viability end point, selenite was approximately 70-fold more cytotoxic than selenate. By contrast, 24 h exposures to either DL- or L-SeMet in the three media caused little or no cytotoxicity. However for 7 d exposures in L15/ex, DL- and L-SeMet were very cytotoxic, even at the lowest tested concentration of 31 μM. By contrast in L15 and L15/FBS, cytotoxicity was only observed with 500 and 1000 μM L-SeMet. In L15/FBS, eelB continued to migrate and form CLS in the presence of SeMet but at 500 μM, cell migration appeared stimulated. As judged from a colony-forming assay over 14 d in L15/FBS, 500 and 1000 μM DL- and L-SeMet inhibited cell proliferation. Overall, the responses of eel cells to selenium depended on the selenium form, concentration, and exposure media, with responses to SeMet being most dependent on exposure media.

摘要

在三种培养基中研究了缺硒和添加硒对美洲鳗鲡脑内皮细胞系(eelB)的影响:完全生长培养基(L15/FBS)、无血清培养基(L15)和基础培养基(L15/ex)。L15/ex仅含有半乳糖和丙酮酸,可用于研究细胞缺硒情况。在没有任何明显硒来源的L15/ex中,eelB细胞存活至少7天,在基质胶上形成毛细血管样结构(CLS),并迁移以愈合伤口。向培养物中添加了三种硒化合物:亚硒酸盐、硒酸盐和硒代蛋氨酸(SeMet)。向eelB细胞培养物中添加亚硒酸盐或硒酸盐24小时会导致细胞活力呈剂量依赖性下降,与暴露培养基无关。尽管随暴露培养基和活力终点而变化,但亚硒酸盐的细胞毒性比硒酸盐高约70倍。相比之下,在三种培养基中24小时暴露于DL-或L-SeMet几乎没有或没有细胞毒性。然而,在L15/ex中暴露7天,DL-和L-SeMet即使在最低测试浓度31μM时也具有很强的细胞毒性。相比之下,在L15和L15/FBS中,仅在500和1000μM L-SeMet时观察到细胞毒性。在L15/FBS中,eelB在存在SeMet的情况下继续迁移并形成CLS,但在500μM时,细胞迁移似乎受到刺激。根据在L15/FBS中14天的集落形成试验判断,500和1000μM DL-和L-SeMet抑制细胞增殖。总体而言,鳗鲡细胞对硒的反应取决于硒的形式、浓度和暴露培养基,对SeMet的反应最依赖于暴露培养基。

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