| Rapid and Simplified Induction of Neural Stem/Progenitor Cells (NSCs/NPCs) and Neurons from Human Induced Pluripotent Stem Cells (hiPSCs)
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Author:
Date:
2021-02-05
[Abstract] Human induced pluripotent stem cells (iPSCs) and their progeny displaying tissue-specific characteristics have paved the way for regenerative medicine and research in various fields such as the elucidation of the pathological mechanism of diseases and the discovery of drug candidates. iPSC-derived neurons are particularly valuable as it is difficult to analyze neural cells obtained from the central nervous system in humans. For neuronal induction with iPSCs, one of the commonly used approaches is the isolation and expansion of neural rosettes, following the formation of embryonic bodies (EBs). However, this process is laborious, inefficient, and requires further purification of the cells. To overcome these limitations, we have developed an efficient neural induction method that allows for ...
[摘要] [摘要]人类诱导的多能干细胞（iPSC）及其后代具有组织特异性，为再生医学的研究铺平了道路，并在疾病的病理机制阐明和候选药物的发现等领域进行了研究。iPSC集-来源的神经元是特别有价值的，因为它是难以分析神经细胞获自人类的中枢神经系统。对于用iPSC诱导神经元，最常用的方法之一是在形成胚体（EB）之后分离和扩展神经玫瑰花结。然而，该过程费力，效率低下，并且需要进一步纯化细胞。为了克服这些限制，我们已经开发出一种高效神经诱导方法，该方法允许来自于7天内的iPSC神经干/祖细胞（NSCs / NPC的）的产生和功能的成熟神经元的。我们的方法产生一个PAX6 -阳性同质细胞群中，皮质神经干细胞/ NPC的，和t他所得的NSCs / NPC的可冷冻保存，膨胀，并分化在功能性成熟神经元。此外，我们的协议将比其他方法便宜，因为该协议在神经诱导期间需要较少的神经补充。本文还介绍了FM1 - 43成像测定法中，其是用于所述的iPSC衍生的突触前评估中有用的人类神经元。该协议为生成NSC / NPC和神经元提供了一种快速且简化的方法，使研究人员能够建立体外细胞模型来研究脑部疾病的病理学。
[背景]人类iPSC于2007年通过使用四种转录因子（Oct4，Sox2，Klf4和c-Myc）对皮肤成纤维细胞进行重编程而首次建立，并且表现出与胚胎干细胞（ESCs）相似的特征，包括其多能性和自我-更新（Takahashi等，2007； ...
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| A Blood-retina Barrier Permeability Assay in Young Mice Using Sulfo-NHS-LC-biotin Perfusion
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Author:
Date:
2018-10-20
[Abstract] Brain and retinal vasculatures exhibit restricted vascular permeability known as blood-brain barrier and blood-retina barrier. Vascular permeability can be evaluated by perfusion of the amine reactive ester derivatives of biotin such as sulfo-NHS-LC-biotin. This protocol describes experimental procedures of sulfo-NHS-LC-biotin perfusion to evaluate retinal vascular permeability. Perfused sulfo-NHS-LC-biotin remained within vessels in wild-type postnatal day 15 (P15) retinas, confirming an intact blood-retina barrier. In contrast, sulfo-NHS-LC-biotin was occasionally detected in extravascular spaces in perfused Eogt−/− retinas suggesting a partly impaired vascular integrity in the absence of Eogt (Sawaguchi et al., 2017).
[摘要] 脑和视网膜脉管系统表现出受限的血管通透性，称为血脑屏障和血 - 视网膜屏障。 血管通透性可以通过灌注生物素的胺反应性酯衍生物如磺基-NHS-LC-生物素来评估。 该方案描述了磺基-NHS-LC-生物素灌注的实验程序，以评估视网膜血管通透性。 灌注的磺基-NHS-LC-生物素保留在野生型出生后第15天（P15）视网膜的血管内，证实了完整的血 - 视网膜屏障。 相比之下，在灌注的 Eogt - / - >视网膜中，偶尔会在血管外空间检测到磺基-NHS-LC-生物素，这表明在没有的情况下血管完整性部分受损。 Eogt >（Sawaguchi et al。>，2017）。
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