{{'Search' | translate}}
 

Sterile Polystyrene Disposable Serological Pipets

公司名称: Fisher Scientific
产品编号: 13-678-11
Bio-protocol()
Company-protocol()
Other protocol()

Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
Author:
Date:
2018-05-20
[Abstract]  Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual metabolic features. Mitochondria not only play a key role in energy metabolism but also in cell cycle regulation of cells. Therefore, mitochondria have emerged as a potential target for anticancer therapy since they are structurally and functionally ... [摘要]  哺乳动物细胞通过线粒体(氧化磷酸化)和非线粒体(糖酵解)代谢产生ATP。已知癌细胞使用不同的策略重新编程它们的代谢以满足能量和合成代谢需要(Koppenol等人,2011; Zheng,2012)。此外,每个癌症组织都有其自己的个体代谢特征。线粒体不仅在能量代谢中起关键作用,而且在细胞的细胞周期调控中也起关键作用。因此,线粒体作为抗癌治疗的潜在靶标已经出现,因为它们在结构和功能上与其非癌对应物不同(D'Souza等人,2011)。我们详细介绍了利用海马XF24细胞外通量分析仪(图1)测量活细胞中氧耗率(OCR)和细胞外酸化率(ECAR)测量的方案。 Seahorse XF24细胞外通量分析仪持续测量细胞上清液中的氧浓度和质子流量(Wu等人,2007)。这些测量结果在OCR和ECAR值中转换,并能够直接定量线粒体呼吸和糖酵解。有了这个协议,我们试图评估三种不同癌细胞系的基线粒体功能和线粒体应激反应细胞毒性测试先导化合物甲磺卡西林,以研究其作用机制。将细胞铺在XF24细胞培养板中并保持24小时。在分析之前,将培养基替换为无缓冲的DMEM pH7.4,然后使细胞在非代谢通量分析前使用Seahorse XF在非CO 2孵育器中平衡以允许精确测量Milli-pH单位改变。 ...

High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level
Author:
Date:
2018-05-20
[Abstract]  The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to evaluate high-dimensional transcriptomic and proteomic data (relating to cellular mRNA and protein), there has not been a method to evaluate corresponding high-dimensional functionomic data (relating to cellular functions) from single cells. Here, we present a protocol to quantitatively measure the differentiation potentials of single human ... [摘要]  在单细胞水平进行细胞转录,信号传导和功能调查的能力为以前所未有的决议研究异质人群开启了机会。 尽管已经开发了评估高维转录组学和蛋白质组学数据(与细胞mRNA和蛋白质有关)的方法,但尚未有方法从单个细胞评估相应的高维功能组学数据(与细胞功能有关)。 在这里,我们提出了一种方案来定量测量单个人造血干细胞和祖细胞的分化潜能,然后根据这些测量结果聚集细胞。 细胞电位的高维功能组分析允许细胞功能与相同祖细胞群体内的分子机制相关联。

【背景】单细胞水平的细胞转录,信号传导和功能单细胞测量技术的发展,以及流式细胞仪等先前存在的技术的发展,使得新镜头能够检测复杂的异质群体。这些方法产生大量数据,这可以借助于降维算法来解释,如使用Mpath,Monocole,PCA,Wishbone或扩散图算法在单细胞RNA-Seq上所示的(Paul等, 2016年;参见 et al。,2017),以及使用tSNE或PhenoGraph的CyTOF(Amir et al。,2013; Levine et al 。,2015)。

我们开发了这个协议,以允许在单细胞环境中对造血祖细胞的大规模培养物进行功能分析和随后的降维。在这个协议中,我们描述了在细胞因子的基质细胞培养物中培养人CD34 ...

产品评论