| Production of Phenotypically Uniform Human Cerebral Organoids from Pluripotent Stem Cells
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Author:
Date:
2021-04-20
[Abstract] Recent advances in stem cell technology have allowed researchers to generate 3D cerebral organoids (COs) from human pluripotent stem cells (hPSCs). Indeed, COs have provided an unprecedented opportunity to model the developing human brain in a 3D context, and in turn, are suitable for addressing complex neurological questions by leveraging advancements in genetic engineering, high resolution microscopy, and tissue transcriptomics. However, the use of this model is limited by substantial variations in the overall morphology and cellular composition of organoids derived from the same pluripotent cell line. To address these limitations, we established a robust, high-efficiency protocol for the production of consistent COs by optimizing the initial phase of embryoid body (EB) formation and ...
[摘要] [摘要]在干细胞技术的最新进展已经使研究人员能够产生3D脑类器官由人多能干细胞（（COS）hPSCs ）。事实上，COS提供了一个前所未有的机会，发展人的大脑在3D场景模型，并反过来，适用于通过利用在进步，基因工程，高分辨率显微镜处理复杂的神经系统的问题，并组织转录。然而，在U SE 该模型的模型受到源自同一多能细胞系的类器官的整体形态和细胞组成的实质性变化的限制。为了解决这些限制，我们建立了坚固的，高-通过优化的初始阶段用于生产相一致的COS效率协议胚状体（EB）形成和神经诱导。使用该协议，采购员可以重复地与产生一个均匀的尺寸，形状，以及跨多个批次的细胞组合物。˚F urthermore，类器官的是发展了延长的时间段（3 - 6个月）显示建立的相对成熟的功能，包括电生理学活性的神经元，少突胶质细胞和祖细胞的产生。因此，该平台提供了可用于研究人脑发育和相关疾病的强大实验模型。
图形摘要：
多能干细胞对脑类器官发育的概述
[背景技术]在最新进展在体外从人多能干细胞（衍生3D脑类器官（COS）的发展hPSCs ...
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| 3D Culture Protocol for Testing Gene Knockdown Efficiency and Cell Line Derivation
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Author:
Date:
2018-06-05
[Abstract] Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ignore the true three-dimensional anatomy of solid tumors. Cells cultured in 2D can also undergo cytoskeletal rearrangements and acquire artificial polarity associated with aberrant gene expression (Edmondson et al., 2014). 3D culture systems that better mimic the in vivo situation have been developed recently. 3D in vitro cancer models (tumorspheres) for studying cancer stem cells have gained increased popularity in the field (Weiswald et al., 2015). Systems that use matrix-embedded or encapsulated spheroids, ...
[摘要] 细胞在固体表面生长为单层的传统二维细胞培养仍然代表了药物检测癌症研究的标准方法。然而，在2D培养物中生长的细胞缺乏相关的细胞基质和细胞 - 细胞相互作用，并且忽略实体肿瘤的真实三维解剖结构。在2D中培养的细胞也可经历细胞骨架重排并获得与异常基因表达相关的人造极性（Edmondson等人，2014）。最近开发出更好地模拟体内情况的3D文化系统。用于研究癌症干细胞的3D体外肿瘤模型（肿瘤球体）在该领域已经获得了越来越多的普及（Weiswald等人，2015）。使用基质嵌入或封装的球体，悬滴培养的球体，磁悬浮系统或3D打印方法的系统已经广泛用于研究和新药筛选。在本文中，我们描述了测试shRNA介导的基因沉默对肿瘤球体形成和生长的影响的详细方案。这种方法允许研究人员测试基因敲低对肿瘤起始细胞生长的影响。正如我们实验室所证实的那样，该方案也可用于直接从肿瘤组织中分离3D癌细胞系。
【背景】3D体外肿瘤细胞模型代表了细胞系与体内生长的肿瘤之间的桥接实验方法（Pampaloni等人，2007; ...
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| Sphere Formation (Osteosphere/Sarcopshere) Assay
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Author:
Date:
2012-12-20
[Abstract] Self-renewing cells from adult tissue (such as bone) that represent a progenitor population can be grown in suspension cultures in the presence of defined serum-free medium. Progenitor cells can be identified by this property of anchorage-independent growth in suspension cultures. These spherical clusters of progenitor bone cells growing under non-adherent conditions are called osteospheres. Such progenitor populations often possess characteristics of multipotency and can differentiate into multiple mesenchymal lineages. Cancer cells capable of growing in suspension have also been reported in osteosarcomas, tumors of the bone tissue. These spherical colonies formed from single cells (clonal) in non-adherent conditions are generally considered to represent self-renewing, stem-like cells ...
[摘要] 来自代表祖细胞群体的成体组织（例如骨）的自我更新细胞可以在确定的无血清培养基的存在下在悬浮培养物中生长。 祖细胞可以通过悬浮培养物中锚定非依赖性生长的这种性质来鉴定。 这些在非粘附条件下生长的祖细胞骨细胞的球形簇称为骨干球。 这样的祖细胞群通常具有多能性的特征并且可以分化成多个间充质谱系。 在骨肉瘤，骨组织的肿瘤中也报道了能够在悬浮液中生长的癌细胞。 在非粘附条件下由单细胞（克隆）形成的这些球形菌落通常被认为代表自我更新的干细胞样细胞，并且可以用于其它测定，例如多能性和有限稀释分析（LDA）。
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