| 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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| A 3D Skin Melanoma Spheroid-Based Model to Assess Tumor-Immune Cell Interactions
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Author:
Date:
2020-12-05
[Abstract] Three-dimensional (3D) tumor spheroids have the potential to bridge the gap between two-dimensional (2D) monolayer tumor cell cultures and solid tumors with which they share a significant degree of similarity. However, the progression of solid tumors is often influenced by the dynamic and reciprocal interactions between tumor and immune cells. Here we present a 3D tumor spheroid-based model that might shed new light on understanding the mechanisms of tumor and immune cell interactions. The model first utilizes the hanging drop assay, which serves as one of the simplest methods for generating 3D spheroids and requires no specialized equipment. Next, pre-established spheroids can be co-cultured either directly or indirectly with an immune cell population of interest. Using skin melanoma, we ...
[摘要] [摘要]三维（3D）肿瘤球体具有弥合二维（2D）单层肿瘤细胞培养物与实体瘤之间的差距的潜力，它们之间有着显着的相似性。然而，实体瘤的进展通常受肿瘤与免疫细胞之间的动态相互作用和相互影响的影响。在这里，我们提出了一个基于3D肿瘤球体的模型，该模型可能会为了解肿瘤与免疫细胞相互作用的机制提供新的思路。的该模型首先利用了悬滴法，这是生成3D球体的最简单方法之一，不需要专门的设备。接下来，可以将预先建立的球体与目标免疫细胞群体直接或间接共培养。使用皮肤黑色素瘤，我们提供了该模型的详细说明，这可能对成功治疗策略的开发具有重要意义。
[背景]三维（3D）肿瘤球体是球形的自组装肿瘤细胞聚集体，类似于微转移瘤并复制实体瘤的许多特征。就像在无血管实体瘤的非增生区域一样，球体内部区域的肿瘤细胞通常表现出扰动的基因和蛋白质表达，新陈代谢改变，细胞周期停滞和坏死（Sant and Johnston ，2017）。但是，用于生成3D椭球体的大多数当前可用技术是耗时，困难和昂贵的。一种简单，快速，简便的生成3D球体的方法是使用悬滴法（Foty ...
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| Integration of Human Induced Pluripotent Stem Cell (hiPSC)-Derived Neurons into Rat Brain
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Author:
Date:
2020-09-05
[Abstract] Human neuron transplantation offers novel opportunities for modeling human neurologic diseases and potentially replacement therapies. However, the complex structure of the human cerebral cortex, which is organized in six layers with tightly interconnected excitatory and inhibitory neuronal networks, presents significant challenges for in vivo transplantation techniques to obtain a balanced, functional and homeostatically stable neuronal network. Here, we present a protocol to introduce human induced pluripotent stem cell (hiPSC)-derived neural progenitors to rat brains. Using this approach, hiPSC-derived neurons structurally integrate into the rat forebrain, exhibit electrophysiological characteristics, including firing, excitatory and inhibitory synaptic activity, and establish ...
[摘要] [摘要] 人类神经元移植为建模人类神经系统疾病和潜在的替代疗法提供了新的机会。然而，人脑皮层的复杂结构分为六层，具有紧密互连的兴奋性和抑制性神经元网络，这对体内移植技术获得平衡，功能稳定和稳态稳定的神经元网络提出了重大挑战。在这里，我们提出了一项协议，将人类诱导的多能干细胞（hiPSC ）衍生的神经祖细胞引入大鼠脑。使用这种方法，hiPSC 衍生的神经元在结构上整合到大鼠前脑中，表现出电生理特性，包括放电，兴奋性和抑制性突触活性，并与宿主电路建立神经元连通性。
[背景] 人类大脑皮层是一个复杂的细胞镶嵌体，在不同的皮质层（I-VI）中包含多样化的神经元亚型，可建立轴突输出和树突状输入的特定模式，提供了皮质电路的基本底物（Rakic，2009; Lodato 等等人，2011； Lui 等人，2011）。特别地，兴奋性和抑制性神经传递的平衡对于适当的脑功能是必需的（Turrigiano和Nelson，2004）。人类诱导的多能干细胞（hiPSC ）可以在人类遗传背景下对人类神经系统疾病进行建模（Dolmetsch和Geschwind，2011； Brennand 等，2015； Vera和Studer，2015）。建立体外系统以将hiPSCs ...
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