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Dulbecco’s modified Eagle’s medium (DMEM)/F-12

Company: Gibco
Catalog#: 11330032
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Production of Phenotypically Uniform Human Cerebral Organoids from Pluripotent Stem Cells
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

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[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 ...

Differentiation of Human Induced Pluripotent Stem Cells (hiPSCs) into Osteoclasts
Author:
Date:
2020-12-20
[Abstract]  

Defects in bone resorption by osteoclasts result in numerous rare genetic bone disorders as well as in some common diseases such as osteoporosis or osteopetrosis. The use of hiPSC-differentiated osteoclasts opens new avenues in this research field by providing an unlimited cell source and overcoming obstacles such as unavailability of human specimens and suitable animal models. Generation of hiPSCs is well established but efficient differentiation of hiPSCs into osteoclasts has been challenging. Published hiPSC-osteoclast differentiation protocols use a hiPSC-OP9 co-culture system or hiPSC-derived embryoid bodies (EBs) with multiple cytokines. Our three-stage protocol consists of 1) EB mesoderm differentiation, 2) expansion of myelomonocytic cells and 3) maturation of hiPSC-osteoclasts.

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[摘要]  [摘要]破骨细胞引起的骨吸收缺陷导致许多罕见的遗传性骨疾病以及某些常见的疾病,例如骨质疏松症或骨质疏松症。采用的hiPSC -分化破骨细胞通过提供无限的细胞来源和克服障碍,如人体标本和合适的动物模型的可用性打开了该领域的新途径。hiPSC的生成已被公认,但是将hiPSC高效分化为破骨细胞一直具有挑战性。发布的hiPSC -osteoclast分化协议使用的hiPSC-OP9共培养体系或hiPSC细胞来源的胚状 具有多种细胞因子的机体(EB)。我们的三阶段协议包含:1)中胚层EB分化,2)的扩张骨髓单核细胞和3)的成熟的hiPSC -osteoclasts。我们通过在Nunclon Sphera微孔板上培养Accutase分离的hiPSCs来产生大小均一的EB,并在4天的细胞因子混合物中促进EB中胚层分化。对于第2阶段,将EBs转移至明胶包被的平板中,并用hM -CSF和hIL-3培养,以扩增骨髓单核细胞群。通过与维生素d,补充hTGF β,HM -CSF和hRANKL ,在第2阶段结束时收集的细胞的diff erentiated成成熟破骨细胞(第3阶段)。与其他技术相比,我们的协议不需要共培养系统。诱导EBs分化为中胚层 均匀的方式; 使用较少的细胞因子进行分化;只需要很短的时间就可以使破骨细胞成熟,并产生足够数量的破骨细胞用于后续的分子分析。

图形摘要: ...

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