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Countess I automated cell counter

CountessTM自动细胞计数器

Company: Thermo Fisher Scientific
Catalog#: C10227
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Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle
Author:
Date:
2020-04-05
[Abstract]  Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have been established for the purification and characterization of exosomes. These include ultracentrifugation, ultrafiltration, size exclusion chromatography, immune capture and precipitation using a proprietary polymer. Here, we provide a protocol based on differential ultracentrifugation and sucrose density gradients tailored for the isolation of crude and ultra-pure exosomes from primary fibroblast cultures derived from adult mouse skeletal ... [摘要]  [摘要 ] 外来体是几乎所有细胞分泌的动态纳米囊泡,并存在于所有生物体液中。鉴于它们的异质含量很高,外泌体已牵涉到它们通过影响细胞-细胞和细胞-ECM通讯而发挥的许多生理和病理过程。近年来,已经建立了越来越多的方法外泌体的纯化和表征。其中包括超速离心,超滤,尺寸排阻色谱,免疫捕获和使用专有聚合物的沉淀。在这里,我们提供了基于差分超速离心和蔗糖密度梯度的协议,该协议专门用于从成年小鼠骨骼肌衍生的原代成纤维细胞培养物中分离粗制和超纯外泌体。可以对该协议进行修改和修改,以从各种组织和体液中分离和表征外泌体。


背景 ] ë xosomes是单膜,异质纳米囊泡直径范围从30至150nm,secre 由所有细胞和存在于几乎所有的体液泰德。外泌体中存在的可溶性和膜大分子,mRNA,microRNA的光谱取决于代谢状态以及分泌这些纳米囊泡的细胞的发育起源。由于它们的货物组成,外泌体可以启动接收细胞中的信号传导途径,并参与了发育,免疫和正常组织生理的维持。在神经退行性疾病,纤维化和癌症等疾病条件下,它们被证明可以触发和传播病理刺激(Rackov 等,2018; Gurunathan 等,2019; van de Vlekkert 等,2019)。在这里,我们描述了从成年小鼠腓肠肌(GA)肌肉建立的成纤维细胞培养物中纯化外泌体的方案(van de Vlekkert ...

Efficient Production of Functional Human NKT Cells from Induced Pluripotent Stem Cells − Reprogramming of Human Vα24+iNKT Cells
Author:
Date:
2017-05-20
[Abstract]  Antigen-specific T cell-derived induced pluripotent stem cells (iPSCs) have been shown to re-differentiate into functional T cells and thus provide a potential source of T cells that could be useful for cancer immunotherapy. Human Vα24+ invariant natural killer T (Vα24+iNKT) cells are subset of T cells that are characterized by the expression of an invariant Vα24-Jα18 paired with Vβ11, that recognize glycolipids, such as α-galactosylceramide (α-GalCer), presented by the MHC class I-like molecule CD1d. Vα24+iNKT cells capable of producing IFN-γ are reported to augment anti-tumor responses, which affects both NK cells and CD8+ cytotoxic T lymphocytes to eliminate MHC- and MHC+ tumor cells, respectively. Here we describe a ... [摘要]  抗原特异性T细胞来源的诱导多能干细胞(iPSCs)已显示重新分化为功能性T细胞,从而提供可用于癌症免疫治疗的T细胞的潜在来源。不变性自然杀伤T(Vα24 + iNKT)细胞的人Vα24 + 细胞是T细胞的子集,其特征在于与Vβ11配对的不变Vα24-Jα18的表达,其识别糖脂,如α-半乳糖神经酰胺(α-GalCer),由MHC I类分子CD1d呈递。据报道能够产生IFN-γ的Vα24 + i / KT细胞增加抗肿瘤反应,其影响NK细胞和CD8 +细胞毒性T淋巴细胞以消除MHC - 和MHC + 肿瘤细胞。在这里,我们描述了将人Vα24 + iNKT细胞重编程到iPSC中的鲁棒方案,然后将其重新分化为Vα24 + iNKT细胞(iPS-Vα24功能的iNKT)。我们进一步提供了测定iPS-Vα24 + iNKT细胞活性的方案。背景 以前有报道说,针对晚期非小细胞肺癌(NSCLC)和头颈部癌症的Vα24 + iNKT细胞癌免疫治疗的临床试验显示疗效,耐受性良好(Motohashi et al。等人,2009; Yamasaki等人,2011)。然而,已知来自外周血单核细胞(PBMC)的Vα24 ...

Determining the Influence of Small Molecules on Hypoxic Prostate Cancer Cell (DU-145) Viability Using Automated Cell Counting and a Cell Harvesting Protocol
Author:
Date:
2016-11-20
[Abstract]  Cell viability assays are an essential aspect of most cancer studies, however they usually require a considerable labor and time input. Here, instead of using the conventional microscopy and hemocytometer cell counting approach, we developed a cell harvesting protocol and combined it with the automated Countess Automated Cell Counter to generate cell viability data. We investigated the effects of dihydroxylated bile acids on the cell viability of prostate cancer cells grown under hypoxic conditions. We observed that for all conditions, cell viability was relatively unchanged, suggesting these molecules had little or no impact on cell viability. The combination of the automated approach and the cell harvesting protocol means this assay is i) easy to perform, ii) extremely reproducible and ... [摘要]  细胞活力测定是大多数癌症研究的一个重要方面,然而,它们通常需要相当多的劳动和时间投入。在这里,而不是使用常规的显微镜和血细胞计数细胞计数方法,我们开发了一个细胞收获协议,并结合自动Countess自动细胞计数器生成细胞活力数据。我们调查二羟基化胆汁酸对缺氧条件下生长的前列腺癌细胞的细胞活力的影响。我们观察到,对于所有条件,细胞活力相对不变,表明这些分子对细胞活力几乎没有或没有影响。自动化方法和细胞收获方案的组合意味着该测定i)容易实施,ii)极其可重复,和iii)其补充更常规的癌症测定数据例如入侵,迁移和粘附。

[背景] 确定任何生物分子的治疗效用是开发新的分子治疗以抵抗癌症进展和发展的关键步骤。作为体外表征的初步步骤,必须评估分子作为抗癌治疗剂的适合性。作为该评估的一部分,细胞活力是小分子的细胞反应的关键决定因素,因为其反映分子维持阈值细胞活力的能力,同时靶向关键癌症进展机制例如。 ,集落性,侵袭和粘附。常规的生存力测定需要包括显微镜,血细胞计数器和手动细胞计数器的大量劳动输入。在这里,我们开发了快速和准确生成细胞活力数据的协议,将补充癌症研究研究(Phelan等人,2016)。

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