Quantification of cell energetics in human subcutaneous adipose progenitor cells after target gene knockdown.
Quantification of cell energetics in human subcutaneous adipose progenitor cells after target gene knockdown.
复制标题
靶基因敲除后人皮下脂肪祖细胞中细胞能量学的定量。
DOI：
10.1016/j.xpro.2023.102607
复制
发表时间：
2023-12-15
期刊：
影响因子：
--
通讯作者：
Feldman, Brian J.
中科院分区：
文献类型：
--
作者：
Li, Liang;Gunewardena, Anya M.;Nyima, Tenzin;Feldman, Brian J.
前脂肪细胞是皮下脂肪组织中的脂肪细胞前体细胞，在人类脂肪组织中保存，具有独特的细胞能量学特征。在这里，我们详细介绍了一种从脂肪组织中获取的原代人类细胞的细胞耗氧率的量化方案。我们描述了原代细胞扩增、细胞接种、转染、分化和呼吸测量的步骤，随后是安捷伦赛马分析公司。生物能量谱的测量和由此产生的数据进一步扩展了我们对从脂肪组织中分离的原代细胞的功能特性的了解。有关本协议使用和执行的完整细节，请参考Chen等人的意见。(2023年)。人皮下前脂肪细胞的培养和分化人皮下前脂肪细胞中特定基因的有效敲除在脂肪生成过程中特定基因对细胞新陈代谢的影响分析出版者注：进行任何实验方案都需要遵守当地实验室安全和伦理的机构指南。前脂肪细胞是皮下脂肪组织中的脂肪细胞前体细胞，在人类脂肪组织中保存，具有独特的细胞能量学特征。在这里，我们详细介绍了一种从脂肪组织中获取的原代人类细胞的细胞耗氧率的量化方案。我们描述了原代细胞扩增、细胞接种、转染、分化和呼吸测量的步骤，随后是安捷伦赛马分析公司。生物能量谱的测量和由此产生的数据进一步扩展了我们对从脂肪组织中分离的原代细胞的功能特性的了解。
Pro-preadipocytes are adipocyte progenitor cells within subcutaneous adipose tissue that are conserved in human adipose tissue with distinct cellular energetics. Here, we detail a protocol to quantify cellular oxygen consumption rates of primary human cells harvested from adipose tissue. We describe steps for primary cell expansion, cell seeding, transfection, differentiation, and respirometry followed by Agilent Seahorse Analytics. The measurement of bioenergetic profiles and resulting data further expand our knowledge of the functional properties of primary cells isolated from adipose tissue. For complete details on the use and execution of this protocol, please refer to Chen et al. (2023). Culture and differentiation of human subcutaneous preadipocytes Effective knockdown of a specific gene in human subcutaneous preadipocytes Analysis of the effect of a specific gene on cellular metabolism during adipogenesis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Pro-preadipocytes are adipocyte progenitor cells within subcutaneous adipose tissue that are conserved in human adipose tissue with distinct cellular energetics. Here, we detail a protocol to quantify cellular oxygen consumption rates of primary human cells harvested from adipose tissue. We describe steps for primary cell expansion, cell seeding, transfection, differentiation, and respirometry followed by Agilent Seahorse Analytics. The measurement of bioenergetic profiles and resulting data further expand our knowledge of the functional properties of primary cells isolated from adipose tissue.
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
--
作者：
Oeckl J;Bast-Habersbrunner A;Fromme T;Klingenspor M;Li Y
通讯作者：
Li Y
影响因子：
16.8
作者：
Symersky, Jindrich;Pagadala, Vijayakanth;Osowski, Daniel;Krah, Alexander;Meier, Thomas;Faraldo-Gomez, Jose D.;Mueller, David M.
通讯作者：
Mueller, David M.