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Trypan Blue solution

台盼蓝溶液

Company: Sigma-Aldrich
Catalog#: T8154
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Isolation and Maintenance of Striatal Neurons
Author:
Date:
2018-04-20
[Abstract]  Primary cultures of murine striatal neurons are widely used to explore cellular mechanisms in neurobiology, including brain diseases. Here we describe a detailed and standardized protocol to dissect and culture embryonic murine striatal neurons GABA-positive/DARPP-32-positive for 12 days in vitro, when they show good neuronal cell connectivity and the presence of dendritic spines, which reflects the maturation of the network. [摘要]  小鼠纹状体神经元的原代培养物被广泛用于探索神经生物学中的细胞机制,包括脑部疾病。 在这里,我们描述了一个详细和标准化的协议解剖和文化胚胎小鼠纹状体神经元GABA阳性/ DARPP-32阳性12天体外,当他们显示良好的神经元细胞连接性和存在的树突 刺,反映了网络的成熟。

【背景】纹状体是运动和奖励系统的重要组成部分,纹状体神经元的功能障碍可导致各种神经元疾病,从强迫性行为(Welch et al。,2007)到神经退行性疾病,如在亨廷顿病中观察到的(Reiner等,1988)。因此,一个良好建立的纹状体文化可能是一个很有价值的研究这些和其他条件的模型。成年纹状体中的主要神经元亚型是中型多刺投射神经元(MSNs),其占所有纹状体神经元的约95%并使用抑制性递质γ-氨基丁酸(GABA)。它们具有中等大小的细胞体,复杂的树突状乔木和高密度的树突棘,可同时接受谷氨酸能和多巴胺输入。其余5%的神经元由GABA能神经间神经元组成(Graveland和DiFiglia,1985; ...

Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
Author:
Date:
2018-02-20
[Abstract]  Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal tissue which is easily attainable, and a promising source of stem cells with no associated ethical concerns. MSCs have been isolated from different regions of the umbilical cord and Wharton’s jelly (WJ) is the gelatinous matrix that surrounds and provides protection to the umbilical cord blood vessels. Being more primitive, MSCs from human umbilical cord exhibit greater proliferative capacity and immunosuppressive ability as compared to adult stem cells which gives them a therapeutic advantage. To meet the requirements for cell therapy, it is important to generate MSCs at a clinical scale by ... [摘要]  间充质干细胞(mesenchymal stem cells,MSCs)目前被认为是“医药信号传导细胞”,在基于细胞的再生治疗方面是一种很有前景的资源。脐带是人类的围产期组织,很容易实现,是一种有前途的干细胞来源,没有相关的伦理问题。 MSC已经从脐带的不同区域分离出来,而Wharton's果冻(WJ)是包围并提供对脐带血管的保护的凝胶状基质。更原始的是,与成人干细胞相比,来自人脐带的MSC表现出更大的增殖能力和免疫抑制能力,这使其具有治疗优势。为了满足细胞疗法的要求,通过遵循不耗时或劳动强度的步骤来产生临床规模的MSC是重要的。在此我们提出了一种简单,高效的方法,通过外植体培养方法从人脐带WJ中分离出MSC,这种方法具有重现性和成本效益。

【背景】间充质干细胞(MSC)具有显着的临床潜力来治疗各种衰弱性疾病,主要是由于其独特的免疫调节作用和再生能力(Caplan and Sorrell,2015)。它们存在于许多组织中(Hass et al。,2011),并被观察到是血管周围的体内(Caplan和Correa,2011)。来源或来源本身的小生境可能导致各种MSC类型之间的重要功能差异(Kwon等人,2016年)。虽然骨髓是研究得最充分和最好的MSCs来源,但也有一定的局限性(Liu et al。,2016)。 ...

In vitro Homeostatic Proliferation of Human CD8 T Cells
Author:
Date:
2017-11-20
[Abstract]  Long-lived T-cell–mediated immunity requires persistence of memory T cells in an antigen-free environment while also maintaining a heightened capacity to recall effector functions. Such antigen-independent homeostatic proliferation is mediated in part by the common gamma-chain cytokines IL-7 and IL-15. To further explore the mechanisms governing maintenance of effector functions in long-lived memory T cells during antigen-independent proliferation, human naïve and memory CD8 T cells can be sorted from peripheral blood mononuclear cells (PBMCs), labeled with the proliferation-tracking dye carboxyfluorescein succinimidyl ester (CFSE), and then purified based on their levels of cell division. This allows investigators to assess differences in the desired molecular target in cells that have ... [摘要]  长寿T细胞介导的免疫需要在无抗原环境中持续存在记忆T细胞,同时还保持提高的回忆效应器功能的能力。这种不依赖于抗原的体内平衡增殖部分由常见的γ链细胞因子IL-7和IL-15介导。为了进一步探索在抗原非依赖性增殖过程中维持长效记忆T细胞中效应子功能的机制,可以从外周血单核细胞(PBMCs)中分选人幼稚和记忆CD8T细胞,用增殖跟踪染料羧基荧光素琥珀酰亚胺酯(CFSE),然后根据它们的细胞分裂水平进行纯化。这使得研究人员能够评估经历细胞因子驱动的增殖的细胞中期望的分子靶点的差异。我们在这里提供了一个协议,用于评估在分裂和未分裂的人类幼稚和记忆CD8 T细胞中的表观遗传程序,在用IL-7和IL-15培养7天之后,以说明这种方法如何阐明控制在长寿记忆CD8T细胞的稳态期间保持效应子功能。
【背景】获得性免疫的一个主要特征是对先前遇到的病原体的免疫记忆的发展(Plotkin等人,2013)。记忆CD8 T细胞在为宿主以前遇到的病原体提供终生保护方面发挥重要作用,但为了提供长寿命的保护,T细胞必须已经获得了在无抗原中持续并保持效应功能的能力环境。在此期间,记忆T细胞响应于IL-7/15细胞因子进行抗原非依赖性增殖,尽管程度不同。

记忆CD8 T细胞的总池是几种不同的细胞亚群的异质组合,其对稳态细胞因子的反应不同。例如,CD8 ...

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