{{'Search' | translate}}
 

Sterile Petri dishes (

细胞培养/培养皿

Company: Thermo Fisher Scientific
Catalog#: 150350
Bio-protocol()
Company-protocol()
Other protocol()

Lentiviral shRNA Screen to Identify Epithelial Integrity Regulating Genes in MCF10A 3D Culture
Author:
Date:
2016-12-05
[Abstract]  MCF10A 3D culture system provides a reductionist model of glandular mammary epithelium which is widely used to study development of glandular architecture, the role of cell polarity and epithelial integrity in control of epithelial cell functions, and mechanisms of breast cancer. Here we describe how to use shRNA screening approach to identify critical cell pathways that couple epithelial structure to individual cell based responses such as cell cycle exit and apoptosis. These studies will help to interrogate genetic changes critical for early breast tumorigenesis. The protocol describes a library of lentiviral shRNA constructs designed to target epithelial integrity and a highly efficient method for lentiviral transduction of suspension MCF10A cultures. Furthermore, protocols are ... [摘要]  MCF10A 3D文化系统提供了腺体乳腺上皮的还原剂模型,其广泛用于研究腺体结构的发育,细胞极性和上皮完整性在上皮细胞功能的控制中的作用以及乳腺癌的机制。在这里我们描述如何使用shRNA筛选方法来识别关键细胞通路,夫妇上皮结构到个别细胞的反应,如细胞周期退出和凋亡。这些研究将有助于询问对早期乳腺肿瘤发生至关重要的遗传变化。该协议描述了设计用于靶向上皮完整性的慢病毒shRNA构建体的文库和用于悬浮MCF10A培养物的慢病毒转导的高效方法。此外,提供的协议设置MCF10A 3D文化在Matrigel的形态和细胞反应研究通过结构化照明和共聚焦显微镜分析免疫染色的三维结构。
关键字: 3D文化,MCF10A,shRNA,上皮完整性,免疫荧光染色,3D成像,形态测量分析

[背景] 上皮细胞形成高度组织的组织结构,其提供物理支持和用于协调细胞信号传导的结构化支架。跨上皮结构的这种协调的信号传导对于上皮生物学是基本的;使得上皮细胞在调节器官大小,形状,功能和基于个体细胞的应答中的动态联合作用(Roignot等人,2013; Shamir和Ewald,2014)。上皮信号传导的联合指挥还提供了一种强有力的肿瘤抑制机制,通过将外部和内部有丝分裂信号门控到静止的上皮组织(Partanen等人,2013; Rejon等人 ...

Preparation of Synovial Mesenchymal Stem Cells from a Rat Knee Joint
Author:
Date:
2016-05-05
[Abstract]  Mesenchymal stem cells (MSCs), first described in human bone marrow, are emerging as promising cell-based therapeutics for a wide range of diseases (Caplan and Correa, 2011). MSCs have been isolated from various organs in the body, and synovial MSCs were first reported by De Bari et al. (2001). We previously reported that synovial MSCs have superior proliferation and chondrogenic potentials as compared to bone marrow-, muscle-, and adipose- derived MSCs in humans (Sakaguchi et al., 2005) and rats (Yoshimura et al., 2007). In addition, administration of synovial MSCs for osteochondral defect promoted cartilage regeneration in a rabbit (Koga et al., 2008) and a pig model (Nakamura et al., 2012). In 2008, we started a clinical trial in human and ... [摘要]  首先在人骨髓中描述的间充质干细胞(MSC)正在作为用于广泛疾病的有希望的基于细胞的治疗剂出现(Caplan和Correa,2011)。 MSC已经从体内各种器官中分离,并且滑膜MSCs首先由De Bari等人(2001)报道。我们先前报道了与人类骨髓,肌肉和脂肪来源的MSC(Sakaguchi等人,2005)和大鼠(Yoshimura等人,2005)相比,滑膜MSC具有更好的增殖和软骨形成潜能> et al。,2007)。此外,施用用于骨软骨缺损的滑膜MSC促进兔子中的软骨再生(Koga等人,2008)和猪模型(Nakamura等人,2012) 。在2008年,我们开始了人类的临床试验,并通过磁共振成像获得令人满意的症状和再生软骨的结果(Sekiya等人,2015)。我们还利用滑膜MSC在大鼠中进行了半月板再生的多个研究线(Horie等人,2009; Horie等人,2012; Katagiri等人al。,2013; Okuno等人,2014; Ozeki等人,2015)。在本文中,我们演示了如何从大鼠膝关节收获包括髌下脂肪垫的滑膜,并描述大鼠滑膜MSC的分离和培养技术。

Comments