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100 mm tissue culture dishes

Corning ® 100mm TC处理培养皿

Company: Corning
Catalog#: 430167
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Generation of Functional Mouse Hippocampal Neurons
Author:
Date:
2020-08-05
[Abstract]  Primary culture of mouse hippocampal neurons is a very useful in vitro model for studying neuronal development, axonal and dendritic morphology, synaptic functions, and many other neuronal features. Here we describe a step-by-step process of generating primary neurons from mouse embryonic hippocampi (E17.5/E18.5). Hippocampal neurons generated with this protocol can be plated in different tissue culture dishes according to different experimental aims and can produce a reliable source of pure and differentiated neurons in less than one week. This protocol covers all the steps necessary for the preparation, culture and characterization of the neuronal culture, including the illustration of dissection instruments, surgical procedure for embryos’ isolation, culturing conditions and ... [摘要]  [摘要] 原代培养小鼠海马神经元是一种非常有用的体外模型用于研究神经元的发育,轴突和树突的形态,突触功能,以及许多其他神经元的特征。这里我们描述了从小鼠胚胎海马(E17.5/E18.5)产生初级神经元的一步一步的过程。根据不同的实验目的,用该方法产生的海马神经元可以在不同的组织培养皿中进行培养,并能在不到一周的时间内产生一个可靠的来源。该方案涵盖了神经元培养物的制备、培养和鉴定的所有必要步骤,包括解剖器械的说明、胚胎分离的手术程序、培养条件以及培养物纯度和分化的评估。通过分析培养6天时的钙显像动力学来评估神经元的活性。

[背景] 海马体是一个非常典型的大脑结构,对重要的大脑功能如记忆、空间导航、情绪记忆和学习至关重要。从解剖学上讲,小鼠海马体有一个清晰的C形结构,很容易定位和分离。在细胞水平上,它主要由锥体细胞组成,与其他脑区相比,中间神经元和胶质细胞较少(Kaech和Banker,2006)。因此,海马体是从野生型或基因工程小鼠模型中产生高纯度原代神经元培养物的理想区域,可用于疾病建模或研究神经元功能的多个方面,如突触传递和电生理特性、对神经毒性的敏感性,分化与衰老(;;;;)。Busche,2018Koyama和Ikegaya,2018Molnar,2011Wu等人,2019Rush等人,2020年

已经制定了许多协议,通过与神经胶质喂食器共同培养神经元来产生皮层和海马神经元(Kaech和Banker,2006),描述了用水凝胶微纤维封装的星形胶质细胞的三维神经元培养系统(Kim等人,2020年),长期向培养基中补充生长因子神经元培养(Ray ...

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 ...

In vivo Tumor Growth and Spontaneous Metastasis Assays Using A549 Lung Cancer Cells
Author:
Date:
2020-04-05
[Abstract]  Metastasis accounts for the majority of cancer related deaths. The genetically engineered mouse (GEM) models and cell line-based subcutaneous and orthotopic mouse xenografts have been developed to study the metastatic process. By using lung cancer cell line A549 as an example, we present a modified protocol to establish the cell line-based xenograft. Our protocol ensures sufficient establishment of the mouse xenografts and allows us to monitor tumor growth and spontaneous metastasis. This protocol could be adapted to other types of established cancer cell lines or primary cancer cells to study the mechanism of metastatic process as well as to test the effect of the potential anti-cancer agents on tumor growth and metastatic capacity. [摘要]  [摘要 ] 中号etastasis应收大多数癌症相关死亡。已经开发了基因工程小鼠(GEM)模型以及基于细胞系的皮下和原位小鼠异种移植物,以研究转移过程。以肺癌细胞系A549为例,我们提出了一种改进的方案来建立基于细胞系的异种移植物。我们的协议可确保小鼠异种移植物的充分建立,并允许我们监测肿瘤的生长和自发转移。该方案可适用于其他类型的已建立癌细胞系或原发性癌细胞,以研究转移过程的机制以及测试潜在抗癌药对肿瘤生长和转移能力的影响。

[背景 ] 的5年生存率速率为患者与晚期阶段肺癌症是少大于15%时,与在大多数的患者死于从转移性疾病(斯特和恩格尔曼,2012 ; 惠特塞特,等人,2013 ; D'安东尼奥等人。,2014 ; Steeg的,2016) 。因此,使用鼠标模型来调查的机制的肺肿瘤侵袭和转移可能促进了发展的新的战略来控制的转移过程。

转移是一个多步骤过程是包括本地侵袭,血管内,并且生存在的循环,外渗,一个d建立macrometastasis 在遥远的站点。小鼠异种移植和基因工程小鼠(GEM)模型均已用于研究肺癌转移。GEM模型模仿了人类肺癌的遗传标记,例如TP53 ,K- ras 和LKB1,为研究转移过程的机制以及研究新型抗癌剂如何影响转移过程提供了免疫系统(Dutt和Wong ,2006年)。 ; Zheng ...

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