Generation of Human Mesenchymal Stem Cell 3D Spheroids Using Low-binding Plates
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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 ...
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Generation of Tumour-stroma Minispheroids for Drug Efficacy Testing
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Author:
Date:
2017-01-05
[Abstract] The three-dimensional organisation of cells in a tissue and their interaction with adjacent cells and extracellular matrix is a key determinant of cellular responses, including how tumour cells respond to stress conditions or therapeutic drugs (Elliott and Yuan, 2011). In vivo, tumour cells are embedded in a stroma formed primarily by fibroblasts that produce an extracellular matrix and enwoven with blood vessels. The 3D mixed cell type spheroid model described here incorporates these key features of the tissue microenvironment that in vivo tumours exist in; namely the three-dimensional organisation, the most abundant stromal cell types (fibroblasts and endothelial cells), and extracellular matrix. This method combined with confocal microscopy can be a powerful tool to ...
[摘要] 组织中细胞的三维组织及其与相邻细胞和细胞外基质的相互作用是细胞反应的关键决定因素,包括肿瘤细胞如何对应激条件或治疗药物的反应(Elliott和Yuan,2011)。在体内,肿瘤细胞被包埋在主要由成纤维细胞形成的基质中,所述成纤维细胞产生细胞外基质并用血管编织。这里描述的3D混合细胞类型球体模型包括了体内存在肿瘤的组织微环境的这些关键特征;即三维组织,最丰富的基质细胞类型(成纤维细胞和内皮细胞)和细胞外基质。该方法结合共聚焦显微镜可以成为不同肿瘤类型的药物敏感性,血管生成和细胞迁移/侵袭测定的有力工具。
背景 传统的单层细胞培养(二维)强化人造环境,其与体内存在的组织细胞大不相同。最重要的区别之一是在单层培养物中,细胞是极化的,即,面向培养物的细胞表面和暴露于培养基的上细胞表面完全不同,经常反对的信号(Fitzgerald等人,2015)。为了解决细胞极化的问题,肿瘤球状体培养越来越多地用于癌症研究。肿瘤球体可以通过减少其通常发生在单层培养物中的生长培养基的扩散和稀释,通过细胞因子和趋化因子复制存在于组织中的三维细胞 - 细胞相互作用和一定程度的旁分泌信号传导(Lawlor等,2002; ...
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3D Gel Invasion Assay of Gastric Cancer Cells with Fibroblasts
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Author:
Date:
2016-05-05
[Abstract] Cancer tissue is composed of cancer cells and a large number of stromal cells including fibroblasts. In order to understand the relationship between fibroblasts and cancer cells during invasion of the stroma, 3D gel invasion assay is useful. Most tumors are associated with a biologically active type of fibroblasts known as cancer-associated fibroblasts (CAFs), which promote the invasion of cancer cells. Here, we describe the method of imaging the invasion by fluorescently labeled CAFs and gastric cancer cells in gels containing extracellular matrix. For two-color fluorescence labeling of living cells, long-chain dialkylcarbocyanines, DiO and DiI were used. This method is also applicable for studying invasion by other stromal cells and cancer cells, and for evaluation of drugs targeting ...
[摘要] 癌组织由癌细胞和大量包括成纤维细胞的基质细胞组成。 为了理解在基质侵入期间成纤维细胞和癌细胞之间的关系,3D凝胶入侵测定是有用的。 大多数肿瘤与称为癌症相关成纤维细胞(CAF)的生物活性类型的成纤维细胞相关,其促进癌细胞的侵入。 在这里,我们描述的成像入侵的荧光标记CAF和胃癌细胞在含有细胞外基质的凝胶中的方法。 对于活细胞的双色荧光标记,使用长链二烷基碳菁,DiO和DiI。 该方法也适用于研究其他基质细胞和癌细胞的侵袭,以及用于评价靶向癌症基质细胞的药物。
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