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Trypan Blue solution

台盼蓝溶液

Company: Sigma-Aldrich
Catalog#: T8154
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Flow Cytometry of CD14, VDR, Cyp27 and Cyp24 and TLR4 in U937 Cells
Author:
Date:
2020-08-05
[Abstract]  Chronic Kidney Disease (CKD) patients present a micro inflammation state due to failure renal function. The calcitriol has been described as an anti-inflammatory factor that might modulates the inflammatory response in CKD patients. However, these patients have deficiency of Calcitriol due to failure renal function. But, synthesis of this vitamin has been reported in extra renal production, as in monocytes. In this context, it has been reported that the supplementation with 25 vitamin D (calcidiol or inactive form of vitamin D) induces monocytes to downregulate inflammation, due to the intracellular 1α-hidroxilase that converts calcidiol to calcitriol in these cells. Besides some reports used RT-qPCR, Western Blot or immunofluorescence techniques to investigate the expression of ... [摘要]  [摘要] 慢性肾脏病(CKD)患者由于肾功能衰竭而出现微炎症状态。骨化三醇被认为是一种抗炎因子,可能调节CKD患者的炎症反应。然而,这些患者由于肾功能衰竭而出现骨化三醇缺乏症。但是,据报道,这种维生素的合成是在肾外产生的,比如在单核细胞中。在这种情况下,有报道称,补充25维生素D(钙二醇或不活跃形式的维生素D)可诱导单核细胞下调炎症反应,这是由于细胞内的1α-羟脯氨酸酶将钙二醇转化为骨化三醇。本研究除采用RT-qPCR、Western-Blot或免疫荧光技术研究炎症和维生素D机械生物标志物在几种疾病中的表达外,本研究还应用流式细胞术技术评价25种维生素D对CD14、Toll样受体4(TLR4)、维生素D受体(VDR)的影响,单核细胞系(U937)中的1-α羟化酶(CYP27)、24-羟化酶(CYP24)。将U937培养物与健康或CKD血清孵育,用或不加25维生素D(50 ng/ml,24 h)处理,以评估CD14、TRL4、VDR、CYP27和CYP24的表达。该方案显示了研究25维生素D处理对细胞内和细胞膜生物标志物表达的影响的优势。此外,与RT-qPCR、westernblot或免疫荧光法相比,该技术不费力,但易于操作和解释。

[背景] ...

Generation of Human Mesenchymal Stem Cell 3D Spheroids Using Low-binding Plates
Author:
Date:
2018-08-20
[Abstract]  The 3D culture of human mesenchymal stem cells (hMSCs) represents a more physiological environment than classical 2D culture and has been used to enhance the MSC secretome or extend cell survival after transplantation. Here we describe a simple and affordable method to generate 3D spheroids of hMSCs by seeding them at high density in a low-binding 96-well plate.

Spheroids of hMSCs cultured in low-binding 96-well plates can be used to study the basic biology of the cells and to generate conditioned media or spheroids to be used in transplantation therapeutic approaches. These MSCs or their secretome can be used as a regenerative therapy and for tissue repair across multiple disease areas, including neurodegeneration.

In comparison to other methods (hanging drop, use ...
[摘要]  人间充质干细胞(hMSC)的3D培养代表比经典2D培养更生理的环境,并且已经用于增强MSC分泌组或移植后延长细胞存活。 在这里,我们描述了一种简单且经济实惠的方法,通过在低密度96孔板中高密度接种hMSC来生成三维球状体。

在低结合96孔板中培养的hMSC的球状体可用于研究细胞的基本生物学并产生用于移植治疗方法的条件培养基或球状体。 这些MSC或其分泌蛋白组可用作再生疗法和用于多个疾病区域的组织修复,包括神经变性。

与其他方法(悬滴,使用凝胶或生物材料,磁悬浮,等)相比,此处描述的方法简单且经济实惠,无需使用专用设备,昂贵材料或复杂试剂。

【背景】 间充质干细胞(MSCs)是开发针对中风或肌萎缩侧索硬化等疾病的新型再生疗法的有吸引力的候选者(Chen et al。,2001; Bang et al。,2005; Boido et al。,2014)。它们的多功能性使得技术的优化和标准化对于确保MSC疗法可以提供尽可能多的益处是必不可少的。使MSC的治疗潜力(例如,增强的抗炎介质分泌)最大化的一种可能方式是以3D方式培养它们(Bartosh ...

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; ...

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