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Penicillin-Streptomycin

青霉素 - 链霉素

Company: Sigma-Aldrich
Catalog#: P0781
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Zebrafish Embryo Xenograft and Metastasis Assay
Author:
Date:
2018-09-20
[Abstract]  Xenograft models, and in particular the mouse xenograft model, where human cancer cells are transplanted into immunocompromised mice, have been used extensively in cancer studies. Although these models have contributed enormously to our understanding of cancer biology, the zebrafish xenograft model offers several advantages over the mouse model. Zebrafish embryos can be easily cultured in large quantities, are small and easy to handle, making it possible to use a high number of embryos for each experimental condition. Young embryos lack an efficient immune system. Therefore the injected cancer cells are not rejected, and the formation of primary tumors and micrometastases is rapid. Transparency of the embryos enables imaging of primary tumors and metastases in an intact and living embryo. ... [摘要]  异种移植模型,特别是小鼠异种移植模型,其中人癌细胞被移植到免疫受损小鼠中,已广泛用于癌症研究。尽管这些模型对我们对癌症生物学的理解做出了巨大贡献,但斑马鱼异种移植模型与鼠标模型相比具有多种优势。斑马鱼胚胎可以很容易地大量培养,体积小且易于处理,因此可以在每种实验条件下使用大量胚胎。年轻的胚胎缺乏有效的免疫系统。因此,注射的癌细胞不被排斥,并且原发性肿瘤和微转移的形成是快速的。胚胎的透明度使得能够在完整和活的胚胎中对原发性肿瘤和转移瘤进行成像。在这里,我们描述了一种方法,其中表达GFP的肿瘤细胞被注射到斑马鱼胚胎的心包空间。在注射后4天,对胚胎成像并分析原发性肿瘤和远端微转移的形成。

【背景】 斑马鱼( Danio rerio )是一种小型淡水鱼,不仅在发育生物学中,而且在生物医学中越来越受欢迎。快速发展的领域之一是使用斑马鱼作为癌症生物学研究的模型。斑马鱼高效可靠地交配,可以产生大量小,透明,外部发育的后代,可以在例如,多孔板中轻松培养(White et al。 >,2013)。有趣的是,人类肿瘤细胞可以植入斑马鱼胚胎中,许多异种移植物能够通过在胚胎中生长,侵入和转移来忠实地重演它们的恶性行为(Lee et al。,2005; Nicoli et al。,2007; Chapman et ...

Generation of Human Mesenchymal Stem Cell 3D Spheroids Using Low-binding Plates
Author:
Date:
2018-08-20
[Abstract]  The 3D culture of human mesenchymal stem cells (hMSCs) represents a more physiological environment than classical 2D culture and has been used to enhance the MSC secretome or extend cell survival after transplantation. Here we describe a simple and affordable method to generate 3D spheroids of hMSCs by seeding them at high density in a low-binding 96-well plate.

Spheroids of hMSCs cultured in low-binding 96-well plates can be used to study the basic biology of the cells and to generate conditioned media or spheroids to be used in transplantation therapeutic approaches. These MSCs or their secretome can be used as a regenerative therapy and for tissue repair across multiple disease areas, including neurodegeneration.

In comparison to other methods (hanging drop, use ...
[摘要]  人间充质干细胞(hMSC)的3D培养代表比经典2D培养更生理的环境,并且已经用于增强MSC分泌组或移植后延长细胞存活。 在这里,我们描述了一种简单且经济实惠的方法,通过在低密度96孔板中高密度接种hMSC来生成三维球状体。

在低结合96孔板中培养的hMSC的球状体可用于研究细胞的基本生物学并产生用于移植治疗方法的条件培养基或球状体。 这些MSC或其分泌蛋白组可用作再生疗法和用于多个疾病区域的组织修复,包括神经变性。

与其他方法(悬滴,使用凝胶或生物材料,磁悬浮,等)相比,此处描述的方法简单且经济实惠,无需使用专用设备,昂贵材料或复杂试剂。

【背景】 间充质干细胞(MSCs)是开发针对中风或肌萎缩侧索硬化等疾病的新型再生疗法的有吸引力的候选者(Chen et al。,2001; Bang et al。,2005; Boido et al。,2014)。它们的多功能性使得技术的优化和标准化对于确保MSC疗法可以提供尽可能多的益处是必不可少的。使MSC的治疗潜力(例如,增强的抗炎介质分泌)最大化的一种可能方式是以3D方式培养它们(Bartosh ...

Isolation, BODIPY Labeling and Uptake of Exosomes in Hepatic Stellate Cells
Author:
Date:
2017-12-05
[Abstract]  Exosomes have emerged as an important mediator of intercellular communication. They are present in extracellular milieu and therefore, easily accessible by neighboring or distant cells. They carry mRNA, microRNAs and proteins within their vesicles and once internalized by recipient cells; they can modulate multiple signaling pathways with pleiotropic effects from inducing antiviral state to disease progression. We have previously shown that hepatitis C virus (HCV) infected hepatocytes or hepatoma cells harboring genome-length replicon secrete exosomes in culture supernatants. These exosomes are taken up by hepatic stellate cells (HSC) and activate them to induce fibrosis during HCV infection. Here, we describe detailed protocols for exosomes isolation and uptake of BODIPY labeled exosomes ... [摘要]  外来体已经成为细胞间通讯的重要介质。 它们存在于细胞外环境中,因此可以被邻近或远处的细胞轻易获得。 它们在囊泡内携带mRNA,microRNA和蛋白质,并一度被受体细胞内化; 它们可以通过诱导抗病毒状态向疾病进展调节多种信号传导途径,具有多效性。 我们以前已经显示,携带基因组长度复制子的丙型肝炎病毒(HCV)感染的肝细胞或肝癌细胞在培养上清液中分泌外泌体。 这些外来体被肝星状细胞(HSC)摄取,并在HCV感染期间激活它们以诱导纤维化。 在这里,我们描述了外来体分离和肝星状细胞摄取BODIPY标记的外来体的详细的协议。

【背景】外来体是直径为40-150nm的小的膜结合的细胞外囊泡。已在所有体液中鉴定出它们,包括尿,羊水,血清,唾液,母乳,脑脊液,鼻分泌物和组织培养的细胞系的上清液中。外泌体通过转移微小RNA(mRNA),mRNA和蛋白质来介导细胞与细胞之间的通讯。 其可被邻近或远处的细胞摄取,并随后促进受体细胞中的信号传导(Schorey等人,2015)。外泌体富含蛋白质,包括四跨膜蛋白家族成员(CD9,CD81,CD63),热休克蛋白(Hsp60,Hsp70,Hsp90)和多泡体蛋白(annexins,RabGTPases和内体分选复合物,蛋白质)。最近的研究表明,外来体可以影响免疫应答(Li等人,2013; ...

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