{{'Search' | translate}}
 

Centrifuge 5427 R

离心机5427 R

Company: Eppendorf
Catalog#: 5427 R
Bio-protocol()
Company-protocol()
Other protocol()

Primary Cultures from Human GH-secreting or Clinically Non-functioning Pituitary Adenomas
Author:
Date:
2018-04-05
[Abstract]  Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological–based on somatostatin and dopamine agonists–approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary adenomas represented a significant advancement in the knowledge of the mechanisms of their development and in the definition of the determinants of their pharmacological sensitivity. Moreover, recent studies identified also in pituitary adenomas putative tumor stem cells representing, according to the current hypothesis, the real cellular targets to eradicate most malignancies. In this protocol, we describe the procedure to establish primary cultures ... [摘要]  垂体腺瘤是更常见的颅内肿瘤之一,通常用基于生长抑素和多巴胺激动剂手术的手术和药物治疗。 虽然多为良性肿瘤,但侵入性行为的发生常常被检测到,导致预后较差。 来自人类垂体腺瘤的原代培养物的使用代表了对其发育机制的知识以及其药理敏感性决定因素的定义方面的显着进步。 此外,最近的研究也在垂体腺瘤中发现了假定的肿瘤干细胞,根据目前的假设,它代表了根除大多数恶性肿瘤的真实细胞靶标。 在这个协议中,我们描述了从人垂体腺瘤建立原代培养的程序,以及如何选择,体外扩增和表型鉴定推定的垂体腺瘤干细胞。

【背景】垂体腺瘤是最常见的颅内肿瘤之一(高达15%),横断面研究发现每100,000名居民中约有90例发病,其中绝大多数为30岁以上的成年人。大约10%的未经选择的垂体在尸检时进行了检查(即考虑到之前未诊断为垂体疾病的受试者的垂体)( ,Molitch,2017)。尽管通常为良性肿瘤,但垂体腺瘤的处理可因与激素分泌过多相关的临床综合征或发展以治疗抗性,高增殖率,快速复发和绒毛外侵袭为特征的侵袭行为而复杂化(Carreno等人,2017)。成年垂体干细胞的持续存在(Florio,2011)导致垂体腺瘤(以及可能的其他良性瘤形成)的发展可以源自具有干细胞特性(主要是自我更新和分化)的肿瘤细胞的亚群能力),正如已经建立的恶性固体和血液肿瘤一样。

最近的实验证据表明,癌症干细胞(CSC)范例也适用于人和小鼠垂体腺瘤(Donangelo等人,2014; ...

RNA Extraction from Wistar Rat Cochlea for qRT-PCR
Author:
Date:
2017-12-05
[Abstract]  Otology research has developed considerably in recent years and molecular analysis is crucial to identify metabolic pathways and therapeutic targets. However, the structure of the cochlea limits the amount of cell mass, and special care is required for RNA extraction. Studies applying this technique to the cochlea are scarce in the literature, and difficulties in this area exist. In the context, we describe a method for extracting RNA in Wistar rats for qRT-PCR analyses in order to clarify certain aspects and obtain good quality material. [摘要]  近年来,耳科学研究发展迅速,分子分析对鉴定代谢途径和治疗靶点至关重要。 然而,耳蜗的结构限制了细胞的数量,特别注意RNA提取。 在耳蜗中使用这种技术的研究在文献中很少,而且在这方面存在困难。 在此背景下,我们描述了提取Wistar大鼠的RNA进行qRT-PCR分析的方法,以澄清某些方面并获得优质的材料。

【背景】了解涉及毛细胞损伤的分子机制对于保持听力非常重要。 来自mRNA的qRT-PCR用于代谢途径和可能的治疗靶标的研究。 耳科学研究通常使用小型啮齿动物,尽管用保存良好的RNA分离感觉细胞样品用于转录分析是一个重大挑战。 用这些动物进行的研究已经提出了在新生儿耳蜗中提取RNA的有效方法,然而,由于少量mRNA的回收,成年耳蜗呈现局限性。 为了评估获得的RNA的质量,可以进行紫外吸收和琼脂糖凝胶电泳。 在此,我们描述了从成年Wistar大鼠获得mRNA的方法,其随后可用于qRT-PCR测定。

In vitro Engineered DNA-binding Molecule-mediated Chromatin Immunoprecipitation (in vitro enChIP) Using CRISPR Ribonucleoproteins in Combination with Next-generation Sequencing (in vitro enChIP-Seq) for the Identification of Chromosomal Interactions
Author:
Date:
2017-11-20
[Abstract]  We have developed locus-specific chromatin immunoprecipitation (locus-specific ChIP) technologies consisting of insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP). Locus-specific ChIP is a method to isolate a genomic region of interest from cells while it also identifies what binds to this region using mass spectrometry (for protein) or next generation sequencing (for RNA or DNA) as described in Fujita et al. (2016a). Recently, we identified genomic regions that physically interact with a locus using an updated form of enChIP, in vitro enChIP, in combination with NGS (in vitro enChIP-Seq) (Fujita et al., 2017a). Here, we describe a protocol on in vitro enChIP to isolate a target locus for identification of ... [摘要]  我们开发了基因座特异性染色质免疫沉淀(基因座特异性芯片)技术,包括插入ChIP(iChIP)和工程DNA-结合分子介导ChIP(enChIP)。 基因座特异性ChIP是一种从细胞中分离感兴趣的基因组区域的方法,同时它还使用质谱(用于蛋白质)或下一代测序(用于RNA或DNA)鉴定什么与该区域结合,如Fujita等人 (2016a)。 最近,我们使用更新后的enChIP形式,结合NGS( in vitro enChIP-Seq),鉴定了与基因座物理相互作用的基因组区域(Fujimita et al。,2017a)。 在这里,我们描述了一个体外试验的方法,用于分离靶基因座以鉴定与基因座物理相互作用的基因组区域。
【背景】阐明基因组功能强调的分子机制需要鉴定与感兴趣的基因组区域相互作用的分子。为此,我们开发了由插入ChIP(iChIP)和工程化DNA结合分子介导的ChIP(enChIP)组成的基因座特异性染色质免疫沉淀技术(基因座特异性ChIP)技术(Fujita等人 ...

Comments