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Minimum Essential Medium Eagle


Company: Sigma-Aldrich
Catalog#: M0268
Other protocol()

Preparation of Primary Cultures of Embryonic Rat Hippocampal and Cerebrocortical Neurons
[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。

Three Dimensional Spheroid Co-culture Invasion Assay
[Abstract]  The assay was developed to investigate the impact of stromal cells of different types (in our case breast cancer associated fibroblasts stably manipulated to modify expression of genes of interest) on the invasive capacity of epithelial cancer cells (in our case breast cancer cell lines) (Verghese et al., 2013). Typical two dimensional invasion assays do necessarily account for the presence of extracellular matrix that is present around the stromal and tumour cells in vivo and therefore cellular behaviour within these cultures may be non-physiological. This spheroid assay was developed to attempt to replicate more closely the environment that is present around breast cancer stromal and tumour cells in actual tumours (Verghese et al., 2013). Extra cellular ... [摘要]  开发该测定以研究不同类型(在我们的情况下乳腺癌相关的成纤维细胞稳定操纵以修饰目的基因的表达)的基质细胞对上皮癌细胞(在我们的乳腺癌细胞系)的侵袭能力的影响( Verghese ,,2013)。典型的二维侵袭测定确实需要考虑在体内基质和肿瘤细胞周围存在的细胞外基质的存在,因此这些培养物中的细胞行为可能是非生理的。开发这种球状体测定以尝试更密切地复制存在于实际肿瘤中的乳腺癌基质和肿瘤细胞周围的环境(Verghese等人,2013)。包括由胶原IV和胶原I组成的细胞外基质,并且成纤维细胞和上皮细胞被给予发展"生理"相互作用的机会(Verghese等人,2013; Hooper等人, ,2006)。该方法由Nowicki等人(2008)开发,并且在Verghese等人(2013)中使用它来公开数据。