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70 μm Cell strainer

Falcon ®70μmCell Strainer,White

Company: Corning
Catalog#: 352350
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Preparation of Primary Cultures of Embryonic Rat Hippocampal and Cerebrocortical Neurons
Author:
Date:
2017-09-20
[Abstract]  This protocol aims at standardizing the procedure to obtain primary cultures of hippocampal and cerebrocortical neurons for in vitro experiments. Cultures should be prepared from cells isolated during embryonic development when neuronal precursor cells are not yet fully differentiated. This helps increasing the quality and quantity of cells, while offering minimal cell death that often occurs during dissociation of differentiated neurons. Cells plated under the appropriate conditions, either in Petri-dishes or in multi-well plates, will develop and establish synaptic contacts over time since the neuronal culture medium provides the nutrients and trophic factors required for differentiation. In this protocol we describe the methodology for the preparation of both cortical and ... [摘要]  该方案旨在标准化海马和脑皮质神经元原代培养物的体外实验。 培养物应从胚胎发育期间分离的细胞制备,当神经元前体细胞尚未完全分化时。 这有助于提高细胞的质量和数量,同时提供通常在分化的神经元解离期间发生的最小的细胞死亡。 在合适的条件下,在培养皿或多孔板中铺板的细胞将随着时间的推移而发展和建立突触接触,因为神经元培养基提供分化所需的营养和营养因子。 在这个协议中,我们描述了制备皮质和海马神经元培养物的方法。
【背景】本方案描述了使用补充有NeuroCultTM SM1的Neurobasal培养基(Chen等人,2008)的大鼠海马和脑皮层神经元的原代培养物的制备。 NeuroCultTM SM1的组成基于B27补充剂的制剂(Brewer等,1993),但是前者的混合物被发现提高了神经元培养的质量,部分地通过用全转运蛋白替代载脂蛋白转运蛋白 Chen et al。,2008)。 此外,NeuroCultTM SM1的化学成分在原始出版物中有更详细的描述,可以更好地控制实验条件。 用化学确定的培养基制备的神经元培养物的特征在于存在低百分比的星形胶质细胞。 通过添加有丝分裂5-氟-2'-脱氧尿苷的化学抑制剂可以防止维持更长时间的培养物中星形胶质细胞的增殖以允许神经元分化。

Melanoma Stem Cell Sphere Formation Assay
Author:
Date:
2017-04-20
[Abstract]  Self-renewal is the ability of cells to replicate themselves at every cell cycle. Throughout self-renewal in normal tissue homeostasis, stem cell number is maintained constant throughout life. Cancer stem cells (CSCs) share this ability with normal tissue stem cells and the sphere formation assay (SFA) is the gold standard assay to assess stem cells (or cancer stem cells) self-renewal potential in vitro. When single cells are plated at low density in stem cell culture medium, only the cells endowed with self-renewal are able to grow in tridimensional clusters usually named spheres. In the recent years, SFA has also been used also to test the effect of several drugs, chemical and natural compounds or microenviromental components on stem cells self-renewal capacity. Here we will ... [摘要]  自我更新是细胞在每个细胞周期复制自身的能力。在正常组织体内平衡的自我更新过程中,干细胞数量在整个生命中保持不变。癌症干细胞(CSCs)与正常组织干细胞共享这种能力,球形成测定(SFA)是评估干细胞(或癌症干细胞)体外自我更新潜力的金标准测定方法。 。当单细胞在干细胞培养基中以低密度铺板时,仅具有自我更新的细胞能够在通常称为球体的三维簇中生长。近年来,SFA也用于测试几种药物,化学和天然化合物或微环境成分对干细胞自我更新能力的影响。在这里,我们将说明一个详细的方案来评估人类黑色素瘤干细胞的自我更新,作为黑色球体生长。

癌症干细胞(CSCs)首先在急性骨髓瘤白血病(Lapidot et al。,1994)中发现,然后在许多实体瘤中鉴定出包括黑素瘤。 CSCs被定义为具有自我更新和肿瘤起始能力的细胞,能够在体内再生整个肿瘤异质性。可以使用基于表型特征或生物学特性的不同方法从肿瘤块中分离CSCs,然后在体外(自我更新)和体内 (致瘤潜力)。使用细胞表面标志物的组合分离黑素瘤CSCs(Fang等人,2005; Monzani等人,2007; Schatton等人, ,2008; Boiko等人,2010; Boonyaratanakornkit等人,2010)或通过特定干细胞培养基中的培养(Perego等人, ,2010; Santini ...

In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
Author:
Date:
2017-04-05
[Abstract]  In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell wall, making them promising tools for exploring host-pathogen interactions during fungal infection. [摘要]  在本协议中,我们描述了如何在小鼠嗜中性粒细胞与真菌白色念珠菌的真菌菌丝共培养的背景下观察嗜中性粒细胞胞外捕获物(NETs)和真菌细胞壁变化。 这些方案很容易调整,以测试与免疫细胞对真菌和细胞壁的影响有关的各种假设,使其成为探索真菌感染期间宿主病原体相互作用的有前景的工具。

C。白色念珠菌是一种多态性机会性酵母,嗜中性粒细胞是免疫细胞,对于这种和其他真菌病原体的防御至关重要(Brown等,2012; Lionakis和Netea,2013)。 NETs是可以针对病原体部署的潜在防御机制,并且已经表明它们优先针对诸如C的微生物细胞部署。白念珠菌菌丝太大而不能吞噬(Urban et al。,2006; Bruns等人,2010; Branzk等人,,2014; Rohm等人,2014)。 NET已经显示含有许多组分,包括髓过氧化物酶,胞外DNA和瓜氨酸化的组蛋白(Amulic等人,2012; ...

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