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RPMI 1640 medium

RPMI 1640 Medium

Company: Thermo Fisher Scientific
Catalog#: 11875119
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In vitro Differentiation of Human iPSC-derived Cardiovascular Progenitor Cells (iPSC-CVPCs)
Author:
Date:
2020-09-20
[Abstract]  Induced pluripotent stem cell derived cardiovascular progenitor cells (iPSC-CVPCs) provide an unprecedented platform for examining the molecular underpinnings of cardiac development and disease etiology, but also have great potential to play pivotal roles in the future of regenerative medicine and pharmacogenomic studies. Biobanks like iPSCORE ( Stacey et al., 2013; Panopoulos et al., 2017), which contain iPSCs generated from hundreds of genetically and ethnically diverse individuals, are an invaluable resource for conducting these studies. Here, we present an optimized, cost-effective and highly standardized protocol for large-scale derivation of human iPSC-CVPCs using small molecules and purification using metabolic selection. We have successfully applied this protocol ... [摘要]  [摘要 ] 诱导性多能干细胞衍生的心血管祖细胞(iPSC-CVPCs)为检查心脏发育和疾病病因的分子基础提供了前所未有的平台,但在再生医学和药物基因组学的未来中也具有重要作用。像iPSCORE这样的生物库(Stacey 等,2013 ;Panopoulos 等,2017), 其中包含由数百个遗传和种族不同的个体产生的iPSC,是进行这些研究的宝贵资源。在这里,我们为小分子大规模衍生人iPSC-CVPCs和代谢选择纯化提供了一种优化,具有成本效益和高度标准化的方案。我们已经成功地应用了该协议,从154种不同的iPSCORE iPSC品系中获得了iPSC-CVPC,从而获得了大量的高纯度心脏细胞。一个重要的我们的协议的组成部分是Ç ELL Ç onfluency 估计S(ccEstimate ),用于估计当iPSC集单层将达到80%汇合,这是用于发起的iPSC-CVPC推导最佳的时间的自动方法,并且使得协议为易于在具有不同增长率的iPSC系列中使用。此外,我们发现跨iPSC-CVPC的细胞异质性是由于两种截然不同的心脏细胞类型(心肌细胞(CMs)和心外膜衍生细胞(EPDCs))的比例不同导致的,这两种细胞在心脏再生中均具有关键作用。该协议消除了iPSC线到线优化的需要,并且可以轻松地进行调整和扩展,以进行高通量研究或生成大量适用于再生医学应用的细胞。

[背景 ] ...

Identification and Quantitation of Leukocyte Populations in Human Kidney Tissue by Multi-parameter Flow Cytometry
Author:
Date:
2018-08-20
[Abstract]  Inflammatory immune cells play direct pathological roles in cases of acute kidney injury (AKI) and chronic kidney disease (CKD). However, the identification and characterization of distinct populations of leukocytes in human kidney biopsies have been confounded by the limitations of immunohistochemical (IHC)-based techniques used to detect them. This methodology is not amenable to the combinations of multiple markers necessary to unequivocally define discrete immune cell populations. We have developed a multi-parameter, flow cytometric-based approach that addresses the need for panels of cell-specific markers in the identification of immune cell populations, allowing both the accurate detection and quantitation of leukocyte subpopulations from a single, clinical kidney biopsy specimen. In ... [摘要]  炎症性免疫细胞在急性肾损伤(AKI)和慢性肾病(CKD)的病例中发挥直接的病理作用。然而,人肾活组织检查中不同白细胞群的鉴定和表征已被用于检测它们的基于免疫组织化学(IHC)的技术的局限性所混淆。该方法不适用于明确定义离散免疫细胞群所必需的多种标志物的组合。我们开发了一种多参数,基于流式细胞仪的方法,解决了在鉴定免疫细胞群体时需要细胞特异性标记组,从而允许从单个临床肾活检标本中准确检测和定量白细胞亚群。在该方法中,将新鲜的人肾组织解离成单细胞悬浮液,然后进行抗体标记和基于流式细胞术的采集和分析。这种新技术为鉴定和计数人类肾脏疾病中的免疫细胞亚群提供了重大进步,并且是补充临床肾脏活组织检查的传统组织病理学检查的有力平台。

【背景】 肾脏疾病的发病率和患病率正在上升,给全世界的医疗保健社区带来了沉重的负担,并显着影响了患者的生活质量(世界肾脏日:慢性肾病,2015年)。肾脏损害可能由多种损伤引起,包括感染,缺血,毒素,高血压,遗传和代谢紊乱(Imig和Ryan,2013)。对功能性损伤的即时反应是急性肾损伤(AKI),这是一种临床综合征,其特征在于生理上通过肾功能的快速(数小时至数天)降低和组织形态学上通过炎性免疫细胞浸润到肾小管中,组织隔室邻接小管。肾脏(Basile ...

Reporter Assay for Semen-mediated Enhancement of HIV-1 Infection
Author:
Date:
2016-07-20
[Abstract]  Semen contains amyloid fibrils that enhance HIV-1 infection (Münch et al., 2007; Kim et al., 2010; Roan et al., 2011; Arnold et al., 2012; Usmani et al., 2014; Roan et al., 2014). Positively charged semen amyloids capture negatively charged viral particles and increase their attachment rates to the cell surface resulting in enhanced fusion and infection (Roan et al., 2009). Since semen is highly cytotoxic, we developed an assay that allows quantification of the infection enhancing activity of semen while minimizing its cell damaging activity. Here, we describe two protocols that allow the quantification of the infectivity enhancing activity of semen using a reporter cell line (TZM-bl cells) or peripheral blood mononuclear ... [摘要]  精液含有增强HIV-1感染的淀粉样蛋白原纤维(Münch等,2007; Kim等,2010; Roan等,2011; Arnold等,2012; Usmani等,2014; Roan et al。 ,2014)。 带正电的精液淀粉样蛋白捕获带负电的病毒颗粒并增加其对细胞表面的附着率,导致增强的融合和感染(Roan等人,2009)。 由于精液具有高度的细胞毒性,我们开发了一种能够定量精液感染增强活性同时最大限度降低其细胞损伤活性的测定方法。 在这里,我们描述允许使用报道细胞系(TZM-bl细胞)或外周血单核细胞(PBMC)定量精液的感染性增强活性的两种方案。

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