Camilla Krizhanovskii, Hjalti Kristinsson, Andris Elksnis, Xuan Wang, Hamid Gavali, Peter Bergsten, Raphael Scharfmann, Nils Welsh
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Methods - EndoC-βH1 cells were cultured at standard conditions in either DMEM or DMEM/F12 culture medium. Cell death was analyzed using propidium iodide staining and flow cytometry. Insulin release and content was quantified using a human insulin ELISA. Results - We presently observe that substitution of DMEM for a DMEM/Ham's F12 mixture (50%/50% vol/vol) renders the cells sensitive to the apoptotic effects of sodium palmitate and sodium palmitate + high glucose leading to an increased cell death. Supplementation of the DMEM culture medium with linoleic acid partially mimicked the effect of DMEM/F12. Culture of EndoC-βH1 cells in DMEM/F12 resulted also in increased proliferation, ROS production and insulin contents, but markers for metabolic stress, autophagy or amyloid deposits were unaffected. 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Results - We presently observe that substitution of DMEM for a DMEM/Ham's F12 mixture (50%/50% vol/vol) renders the cells sensitive to the apoptotic effects of sodium palmitate and sodium palmitate + high glucose leading to an increased cell death. Supplementation of the DMEM culture medium with linoleic acid partially mimicked the effect of DMEM/F12. Culture of EndoC-βH1 cells in DMEM/F12 resulted also in increased proliferation, ROS production and insulin contents, but markers for metabolic stress, autophagy or amyloid deposits were unaffected. 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引用次数: 22
摘要
目的-已知在体外培养时,人胰岛会因棕榈酸钠的游离脂肪酸而死亡。这与EndoC-βH1细胞形成对比,在我们的研究中,EndoC-βH1细胞对棕榈酸钠诱导细胞死亡的作用不敏感,这使得这些细胞似乎不适合进行脂肪毒性研究。然而,EndoC-βH1细胞通常在基于Dulbecco's Modified Eagle Medium (DMEM)的营养混合物中培养,这可能不是研究脂肪毒性的最佳选择。本研究的目的是确定使内啡肽-βH1细胞对棕榈酸钠的毒性作用敏感的培养条件。方法- EndoC-β h1细胞在标准条件下分别在DMEM或DMEM/F12培养基中培养。采用碘化丙啶染色和流式细胞术分析细胞死亡情况。采用人胰岛素酶联免疫吸附测定胰岛素释放量和含量。结果-我们目前观察到,DMEM取代DMEM/Ham's F12混合物(50%/50% vol/vol)使细胞对棕榈酸钠和棕榈酸钠+高糖的凋亡作用敏感,导致细胞死亡增加。在DMEM培养基中添加亚油酸可以部分模拟DMEM/F12的效果。EndoC-βH1细胞在DMEM/F12中培养也导致增殖、ROS产生和胰岛素含量增加,但代谢应激、自噬或淀粉样蛋白沉积的标志物未受影响。结论-内啡肽-β h1细胞的培养条件可以改变,因此这些细胞在棕榈酸钠的作用下表现出脂肪毒性的迹象。
EndoC-βH1 cells display increased sensitivity to sodium palmitate when cultured in DMEM/F12 medium.
Aims - Human pancreatic islets are known to die in response to the free fatty acid of sodium palmitate when cultured in vitro. This is in contrast to EndoC-βH1 cells, which in our hands are not sensitive to the cell death-inducing effects sodium palmitate, making these cells seemingly unsuitable for lipotoxicity studies. However, the EndoC-βH1 cells are routinely cultured in a nutrient mixture based on Dulbecco's Modified Eagle Medium (DMEM), which may not be the optimal choice for studies dealing with lipotoxicity. The aim of the present investigation was to define culture conditions that render EndoC-βH1 cells sensitive to toxic effects of sodium palmitate. Methods - EndoC-βH1 cells were cultured at standard conditions in either DMEM or DMEM/F12 culture medium. Cell death was analyzed using propidium iodide staining and flow cytometry. Insulin release and content was quantified using a human insulin ELISA. Results - We presently observe that substitution of DMEM for a DMEM/Ham's F12 mixture (50%/50% vol/vol) renders the cells sensitive to the apoptotic effects of sodium palmitate and sodium palmitate + high glucose leading to an increased cell death. Supplementation of the DMEM culture medium with linoleic acid partially mimicked the effect of DMEM/F12. Culture of EndoC-βH1 cells in DMEM/F12 resulted also in increased proliferation, ROS production and insulin contents, but markers for metabolic stress, autophagy or amyloid deposits were unaffected. Conclusions - The culture conditions for EndoC-βH1 cells can be modified so these cells display signs of lipotoxicity in response to sodium palmitate.
期刊介绍:
Islets is the first international, peer-reviewed research journal dedicated to islet biology. Islets publishes high-quality clinical and experimental research into the physiology and pathology of the islets of Langerhans. In addition to original research manuscripts, Islets is the leading source for cutting-edge Perspectives, Reviews and Commentaries.
Our goal is to foster communication and a rapid exchange of information through timely publication of important results using print as well as electronic formats.