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Author:
Date:
2018-06-05
[Abstract] Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ignore the true three-dimensional anatomy of solid tumors. Cells cultured in 2D can also undergo cytoskeletal rearrangements and acquire artificial polarity associated with aberrant gene expression (Edmondson et al., 2014). 3D culture systems that better mimic the in vivo situation have been developed recently. 3D in vitro ...
[摘要] 细胞在固体表面生长为单层的传统二维细胞培养仍然代表了药物检测癌症研究的标准方法。然而，在2D培养物中生长的细胞缺乏相关的细胞基质和细胞 - 细胞相互作用，并且忽略实体肿瘤的真实三维解剖结构。在2D中培养的细胞也可经历细胞骨架重排并获得与异常基因表达相关的人造极性（Edmondson等人，2014）。最近开发出更好地模拟体内情况的3D文化系统。用于研究癌症干细胞的3D体外肿瘤模型（肿瘤球体）在该领域已经获得了越来越多的普及（Weiswald等人，2015）。使用基质嵌入或封装的球体，悬滴培养的球体，磁悬浮系统或3D打印方法的系统已经广泛用于研究和新药筛选。在本文中，我们描述了测试shRNA介导的基因沉默对肿瘤球体形成和生长的影响的详细方案。这种方法允许研究人员测试基因敲低对肿瘤起始细胞生长的影响。正如我们实验室所证实的那样，该方案也可用于直接从肿瘤组织中分离3D癌细胞系。
【背景】3D体外肿瘤细胞模型代表了细胞系与体内生长的肿瘤之间的桥接实验方法（Pampaloni等人，2007; ...
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Author:
Date:
2015-09-05
[Abstract] The plant Golgi apparatus is composed of numerous stacks of cisterna, designated as cis, medial, and trans Golgi cisternae; these stacks move within the cytoplasm along the actin cytoskeleton. Cis cisternae receive secretory products from endoplasmic reticulum (ER) and they subsequently progress through the stack to the trans cisternae, where they are sorted to other destinations, including cell wall, plasma membrane (PM), vacuoles, and chloroplasts. In addition, the plant Golgi apparatus plays a role of glycosylating proteins as well as ...
[摘要] 植物高尔基体包括许多叠状体，称为顺，内侧和反式高尔基这些堆栈在细胞质内沿着肌动蛋白细胞骨架移动。 Ctern 池从内质网（ER）接收分泌产物，并且随后通过堆叠进入反转池，在那里它们被分选到其他目的地，包括细胞壁，血浆膜（PM），液泡和叶绿体。此外，植物高尔基体起到糖基化蛋白质以及合成细胞壁多糖如半纤维素和果胶的作用。该协议描述了使用共聚焦激光显微镜（CLSM），全内反射荧光显微镜（TIRF）和高压的免疫金标记标记定位于拟南芥幼苗的植物高尔基体的荧光标记的蛋白质的程序冷冻/冷冻取代的样品通过透射电子显微镜（TEM）。我们特别关注长期时间推移成像和蛋白质本地化在高尔基内的子域。该方案还可以用于其他细胞器，组织和植物物种。
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