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Fetal bovine serum

胎牛血清

Company: GE Healthcare
Catalog#: SV30160.03
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A Method for Extracting the Nuclear Scaffold from the Chromatin Network
Author:
Date:
2018-04-20
[Abstract]  Each cell contains many large DNA polymers packed in a nucleus of approx. 10 μm in diameter. With histones, these DNA polymers are known to form chromatins. How chromatins further compact in the nucleus is unclear but it inevitably depends on an extensive non-chromatin nuclear scaffold. Imaging of endogenous chromatin network and the complementary scaffold that support this network has not been achieved but biochemical and proteomic investigations of the scaffold can still provide important insights into this chromatin-organizing network. However, this demands highly inclusive and reproducible extraction of the nuclear scaffold. We have recently developed a simple protocol for releasing the scaffold components from chromatins. The inclusiveness of the extract was testified by the ... [摘要]  每个细胞都含有许多大型DNA聚合物,其中包含大约一个核。直径10微米。用组蛋白,已知这些DNA聚合物形成染色质。染色质在核中如何进一步致密还不清楚,但它不可避免地依赖于广泛的非染色质核支架。内源性染色质网络的成像和支持该网络的互补支架尚未实现,但支架的生化和蛋白质组学研究仍然可以提供关于该染色质组织网络的重要见解。但是,这需要高度包容和可重复的提取核支架。我们最近开发了一个简单的协议,用于从染色质中释放脚手架组件。提取物的包容性由以下观察结果证实:当从核中提取时,剩余的核染色质被释放为延伸且通常平行的染色质纤维。基本上,该方案包括纯核的产生,用Triton X-100处理细胞核以产生包膜消耗的细胞核(TxN),并在含蔗糖的缓冲液中在500mM NaCl中提取细胞核。 TxN的这个组合提取被称为TxNE。

【背景】通过蛋白质和核糖核蛋白的复杂支架,染色质在细胞核中密集并动态地压缩。与细胞骨架网络不同(Fischer和Fowler,2015),对这种核支架的显微观察在技术上是具有挑战性的。这可能反映了每个细胞核内染色质的主导地位,支架与细胞核交织在一起。核的球形排列也对成像这种支架结构造成挑战。核支架的主要元素是核层(NL)(Gruenbaum和Foisner,2015)。 ...

Isolation and Expansion of Mesenchymal Stem Cells from Murine Adipose Tissue
Author:
Date:
2017-08-20
[Abstract]  Mesenchymal stem cells (MSCs) are currently intensively studied due to significant promise which they represent for successful implementations of future cell therapy clinical protocols. This in turn emphasizes importance of careful preclinical studies of MSC effects in various murine disease models. The appropriate cell preparations with reproducible biological properties are important to minimize variability of results of experimental cell therapies. We describe here a simple protocol for isolation of murine MSCs from adipose tissues and their reproducible multi-log expansion under hypoxia conditions. [摘要]  间充质干细胞(MSC)目前正在深入研究,因为它们代表未来细胞治疗临床方案的成功实施的重大前景。 这又强调了对各种鼠疾病模型中MSC效应的仔细临床前研究的重要性。 具有可重现的生物学性质的合适的细胞制剂对于最小化实验细胞疗法结果的变异性是重要的。 我们在这里描述了一种用于从脂肪组织中分离鼠MSC的简单方案及其在缺氧条件下的可重复的多对数扩增。
【背景】最初由Friedenstein鉴定的MSC是成纤维细胞样形态的骨髓细胞,粘附于塑料和高自我更新能力,导致体外成纤维细胞样集落的形成(Friedenstein等,1976; Review in Phinney andSensebé ,2013)。 MSCs由于其在医学上的潜在应用,目前是研究最成熟的成体祖细胞类型之一。这些细胞可以从各种器官中分离(Murray等,2014),并且被认为是源于血管,以周细胞或血管壁细胞。除了能够沿着成骨,脂肪形成和软骨形成谱系分化的能力之外,MSC具有免疫调节特性,并且被认为参与对组织损伤的反应,以及通过其影响巨噬细胞极化的能力来组织抗炎反应(Prockop,2013; Caplan,2016)。
   鉴于这些特性,MSCs代表了未来相关细胞治疗临床方案的成功实施的巨大前景。这反过来强调了在各种鼠疾病模型中使用MSC进行仔细临床前研究的重要性。制备大量具有可重复生物学特性的合适细胞样品的能力对于在开发基于MSC的实验细胞疗法期间最小化结果的变异性至关重要。然而,与具有强抗氧化防御性并因此在大气氧条件下相当好的人类MSC不同,小鼠MSC对氧应激更敏感,并且在常规CO2培养箱中培养时具有有限的寿命和扩张能力。相反,在缺氧条件下培养这些细胞,相反,显着延长了它们的寿命,并允许多对数扩增,提供足够量的具有可重复性质的细胞材料,用于用鼠实验疾病模型重复实验(Boregowda等,2012; ...

MHC Class II Tetramer Labeling of Human Primary CD4+ T Cells from HIV Infected Patients
Author:
Date:
2017-03-20
[Abstract]  Major Histocompatibility Complex (MHC) tetramers have been used for two decades to detect, isolate and characterize T cells specific for various pathogens and tumor antigens. In the context of Human Immunodeficiency Virus (HIV) infection, antigen-specific CD8+ T cells have been extensively studied ex vivo, as they can be readily detected by HIV peptide-loaded MHC class I tetramers. In contrast, the detection of HIV-specific CD4+ T cells has proven more challenging, due to the intrinsically lower clonal expansion rates of CD4+ T cells, and to the preferential depletion of HIV-specific CD4+ T cells in the course of HIV infection.

In the following protocol, we describe a simple method that facilitates the identification of CD4+ ...
[摘要]  主要组织相容性复合物(MHC)四聚体已经使用二十年来检测,分离和表征各种病原体和肿瘤抗原特异性的T细胞。在人类免疫缺陷病毒(HIV)感染的背景下,抗原特异性CD8 +细胞已经在体外广泛研究,因为它们可以容易地被HIV肽 - 加载MHC I类四聚体。相比之下,HIV特异性CD4 + sup + T细胞的检测已被证明更具挑战性,因为CD4 + sup + T细胞的本质上较低的克隆扩增率以及优先艾滋病毒感染过程中艾滋病毒特异性CD4 + T细胞的消耗。 在以下协议中,我们描述了一种简单的方法,该方法有助于使用肽负载的MHC II类四聚体来鉴定HIV-1衣壳表位特异性的CD4 + / T细胞。可以分析四聚体标记的CD4 T细胞的细胞表面表型和/或FACS分选用于进一步的下游应用。成功检测特异性CD4 + / / T细胞离体的关键是选择导致高亲和力T细胞受体(TCR)的肽/ MHC II组合, (Benati等人,2016)。 MHC II四聚体阳性细胞的可靠检测的第二个关键点是系统地使用负载无关肽的对照四聚体,样品和对照管在相同的条件下进行处理。背景 在用抗PE微珠对四聚体-PE标记的细胞进行磁力富集后,在纯化的CD4 + T细胞中检测到罕见的HIV特异性MHC II四聚体阳性细胞(Seth等人, em>。,2005)。我们发现使用经验证的肽/ MHC ...

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