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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Long-distance Transport in Bacterial Swarms Revealed by Single Nanoparticle Tracking
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Author:
Date:
2020-11-05
[Abstract] During swarming, high density flagella-driven bacteria migrate collectively in a swirling pattern on wet agar surfaces, immersed in a thin viscous fluid layer called “swarm fluid”. Though the fluid environment has essential role in the emergence of swarming behavior, the microscopic mechanisms of it in mediating the cooperation of bacteria populations are not fully understood. Here, instead of micro-sized tracers used in previous research, we use gold nanorods as single particle tracers to probe the dynamics of the swarm fluid. This protocol includes five major parts: (1) the culture of swarming bacterial colony; (2) the preparations of gold nanorod tracers and the micro-spraying technique which are used to put the nanotracers into the upper fluid of bacterial swarms; (3) ...
[摘要] [摘要]在群体聚集期间,高密度的鞭毛驱动细菌以漩涡状在湿的琼脂表面集体迁移,并浸入称为“群液”的稀薄粘性液层中。尽管流体环境在群居行为的出现中起着至关重要的作用,但其在介导细菌种群合作方面的微观机制尚不完全清楚。在这里,代替以前的研究中使用的微型示踪剂,我们使用金纳米棒作为单颗粒示踪剂来探测群流体的动力学。该方案包括五个主要部分:(1)大量细菌菌落的培养;(2)金纳米棒示踪剂的制备和微喷涂技术被用来放nanotracers进入细菌群的上层液体;(3)成像与跟踪;(四)其他必要的对照实验;(5)数据分析与物理模型拟合。通过这种方法,纳米尺寸的示踪剂可以在运动细胞上方长距离移动,而不会与细菌体直接碰撞。这样,可以以高时空分辨率跟踪群流体的微观动力学。此外,对多粒子轨迹的综合分析提供了流体动力学的系统可视化。该方法有望探测其他天然或人工活性物质系统的流体动力学。
[背景]近来,集体米的细菌otions已经吸引了很多关注的活性物质/流体领域(Rabani等,2013;张,H 。等,2010;马尔凯蒂。等,2013) ...
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Analysis of the Effect of Sphingomyelinase on Rubella Virus Infectivity in Two Cell Lines
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Author:
Date:
2018-09-05
[Abstract] Rubella is a mildly contagious disease characterized by low-grade fever and a morbilliform rash caused by the rubella virus (RuV). Viruses often use cellular phospholipids for infection. We studied the roles of cellular sphingomyelin in RuV infection. Treatment of cells with sphingomyelinase (SMase) inhibited RuV infection in rabbit kidney-derived RK13 cells and African green monkey (Cercopithecus aethiops) kidney-derived Vero cells. Our data further demonstrated that RuV used cellular sphingomyelin and cholesterol for its binding to cells and membrane fusion at the step of virus entry. Detailed protocols of our assays, which assess the effects of SMase treatment on RuV infectivity in RK13 and Vero cells, are described.
[摘要] 风疹是一种轻度传染性疾病,其特征是低风热和由风疹病毒(RuV)引起的麻疹样皮疹。 病毒通常使用细胞磷脂进行感染。 我们研究了细胞鞘磷脂在RuV感染中的作用。 用鞘磷脂酶(SMase)处理细胞抑制兔肾衍生的RK13细胞和非洲绿猴( Cercopithecus aethiops )肾源性Vero细胞中的RuV感染。 我们的数据进一步证明RuV在病毒进入的步骤中使用细胞鞘磷脂和胆固醇来结合细胞和膜融合。 描述了我们的测定的详细方案,其评估了SMase处理对RK13和Vero细胞中RuV感染性的影响。
【背景】风疹病毒(RuV)是一种正链RNA病毒,属于 Togaviridae 家族中的 Rubivirus 属。该家族有两个属, Rubivirus 和 Alphavirus 。风疹病毒是属的鞋底构件的风疹病毒,而许多病毒,例如塞姆利基森林病毒(SFV)和辛德毕斯病毒(SINV),是归类于甲病毒属。 RuV是风疹和先天性风疹综合征(CRS)的致病因子。风疹的特征是低烧,麻疹样皮疹和淋巴结肿大。它通常是一种轻微的疾病。然而,CRS是一种严重的疾病。 CRS导致在怀孕早期患有风疹的母亲所生的新生儿出现多器官缺陷。白内障,感音神经性听力损失和心血管缺陷在CRS中很常见。
以前的研究表明,细胞膜脂质作为RuV感染的结合或进入因子(Mastromarino ...
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