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HyClone Fetal Bovine Serum (FBS)

胎牛血清

Company: GE Healthcare
Catalog#: SV30160.03
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Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors
Author:
Date:
2020-05-20
[Abstract]  Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on antigen cross-presentation, a critical step for inducing CD8+ T cell cytotoxic responses. The rarity of naturally occurring cDC1s and lack of in vitro methodologies for the generation of pure cDC1 populations strongly hinders the study of cDC1 lineage specification and function. Here, we describe a protocol for the generation of induced DCs (iDCs) by lentiviral-mediated expression of the transcription factors PU.1, IRF8 and ... [摘要]  [摘要] 转录因子组合的异位表达最近被证明可以将分化的体细胞重编程为树突状细胞(DC)谱系,而不会回复到多能状态。DC具有诱导有效和持久的适应性免疫应答的能力。在特定的常规类型1的DC(cDC1s)练成抗原交叉呈递,用于诱导CD8的关键步骤+ T细胞的细胞毒性应答。天然存在的cDC1的稀有性和缺乏用于生成纯cDC1群体的体外方法论,严重阻碍了对cDC1谱系规格和功能的研究。在这里,我们描述了用于生成感应DC(iDC)的协议 慢病毒介导的转录因子PU.1,IRF8和BATF3在小鼠胚胎成纤维细胞中的表达。iDC 在9天内获得DC形态,cDC1表型和转录特征。使用此协议生成的iDC 具有对炎症刺激,吞噬死细胞,将抗原加工并交叉呈递给CD8 + T细胞的功能。DC重新编程提供了一个简单易处理的系统,可以生成大量的cDC1类细胞用于高内涵筛选,从而开辟了新途径,可以更好地了解cDC1的规格和功能。将来,在成纤维细胞中忠实诱导cDC1命运可能会导致产生患者特定的疫苗接种DC。

[背景技术树突状细胞(DC)是专业的抗原呈递细胞,专门用于识别,加工和呈递T细胞抗原,在诱导适应性免疫应答和免疫记忆中起关键作用(Me rad 等,2013)。DC可以分为4个主要子集:浆细胞样DC(pDC ),大量1型干扰素的产生者,循环单核细胞衍生的单核细胞衍生DC 和常规DC(cDC ...

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; ...

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