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DPBS, calcium, magnesium

DPBS,钙,镁

Company: Thermo Fisher Scientific
Catalog#: 14040
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Retinal Explant Culture
Author:
Date:
2014-01-20
[Abstract]  A particularly powerful culture method for the retina is the explant assay, which consists in culturing a small piece of retina on an organotypic filter. Retinal explants can be prepared any time between embryonic day 13 (E13) and postnatal day 4 (P4). Although retinal ganglion cells tend to degenerate shortly after they are generated in explants, and photoreceptor cells do not grow extended outer segments, the explants will develop very similarly to a retina in vivo and generate all the different retinal cell types that will migrate to the appropriate layer. The retinal explant culture assay is particularly useful in cases where a mouse mutant is embryonic lethal and its retinal development cannot be studied in vivo. Because retinal explants can be prepared from ... [摘要]  视网膜的特别强大的培养方法是外植体测定,其包括在器官型过滤器上培养小片视网膜。视网膜外植体可以在胚胎第13天(E13)和出生后第4天(P4)之间的任何时间进行。虽然视网膜神经节细胞在外植体中产生后不久就会退化,并且感光细胞不会扩展外延伸的外切片,外植体将在体内与视网膜非常相似,并产生将迁移到适当的所有不同的视网膜细胞类型层。视网膜外植体培养测定在小鼠突变体是胚胎致死性并且其视网膜发育不能在体内研究的情况下特别有用。因为视网膜外植体可以从胚胎动物制备并用病毒载体电穿孔或感染,因此也是胚胎期基因功能研究的有用途径。在这里,我们提出了我们在各种出版物中广泛使用的视网膜外植体培养方法(Kechad等,2012; Cayouette等,2003; Cayouette和Raff,2003; Elliott等,2008)。

Dissociated Retinal Cell Culture
Author:
Date:
2014-01-20
[Abstract]  The retina is a relatively simple and accessible part of the central nervous system, making it a powerful model to study cell fate specification mechanisms. Multipotent retinal progenitor cells (RPCs) give rise to seven major classes of retinal cell types. Mechanisms regulating cell fate choice in the retina depend on both cell intrinsic and environmental factors, but their relative contribution to specific cell fate decisions remains unclear. Dissociated retinal cell cultures provide a great assay to study this problem. RPCs are cultured in serum-free and extract-free medium, providing the investigator with a control over the environment to address questions related to the effects of a particular molecule on the development of retinal neurons. In addition, dissociated cell cultures can ... [摘要]  视网膜是中枢神经系统相对简单易用的部分,是研究细胞命运规范机制的有力模型。多能视网膜祖细胞(RPC)产生七大类视网膜细胞类型。调节视网膜细胞命运选择的机制取决于细胞内在和环境因素,但它们对特定细胞命运决定的相对贡献仍不清楚。分离的视网膜细胞培养物提供了一个很好的分析来研究这个问题。 RPC在无血清和无提取物培养基中培养,为研究者提供对环境的控制,以解决与特定分子对视网膜神经元发育的影响有关的问题。此外,分离的细胞培养物可用于通过从其正常环境中分离RPC来研究细胞内在机制的重要性(Cayouette等人,2003; Jensen和Raff,1997)。下述方法适用于RPC的克隆密度培养。在这样的培养物中,RPC彼此分离并且从后神经元分离。它们分化和分化成不同的视网膜细胞类型以形成小菌落或“克隆”。在最近的一项研究中,我们发现这些克隆与细胞数量和细胞类型组成方面在视网膜原位发生的克隆是无法区分的,这表明细胞内机制在视网膜发育中起关键作用(Cayouette et al。 ,2003)。

Primary Culture of SVZ-derived Progenitors Grown as Neurospheres
Author:
Date:
2013-08-20
[Abstract]  SVZ-derived progenitors grown as neurospheres is a well-known model to study neural stem cell and progenitor functions such as proliferation, differentiation/self-renewal, and migration (Durbec and Rougon, 2001). This protocol is for preparing a culture of SVZ-derived progenitors from 8 early postnatal mouse brains (P0 to P3). One week after cell plating, we can observe round floating neurospheres, each resulting from the clonal expansion of a single EGF/FGF responsive neural progenitor. [摘要]  作为神经球生长的SVZ衍生的祖细胞是研究神经干细胞和祖细胞功能如增殖,分化/自我更新和迁移的众所周知的模型(Durbec和Rougon,2001)。 该方案用于从8只早期出生后的小鼠脑(P0至P3)制备SVZ衍生祖细胞的培养物。 细胞电镀后一周,我们可以观察到圆形浮游神经球,每个由单个EGF / FGF反应性神经祖细胞的克隆扩增产生。

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