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GibcoTM Amphotericin B

两性霉素B

Company: Thermo Fisher Scientific
Catalog#: 15290026
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Heterochronic Pellet Assay to Test Cell-cell Communication in the Mouse Retina
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Date:
2017-02-05
[Abstract]  All seven retinal cell types that make up the mature retina are generated from a common, multipotent pool of retinal progenitor cells (RPCs) (Wallace, 2011). One way that RPCs know when sufficient numbers of particular cell-types have been generated is through negative feedback signals, which are emitted by differentiated cells and must reach threshold levels to block additional differentiation of that cell type. A key assay to assess whether negative feedback signals are emitted by differentiated cells is a heterochronic pellet assay in which early stage RPCs are dissociated and labeled with BrdU, then mixed with a 20-fold excess of dissociated differentiated cells. The combined cells are then re-aggregated and cultured as a pellet on a membrane for 7-10 days in vitro. During ... [摘要]  构成成熟视网膜的所有七种视网膜细胞类型都是由普通的多能视网膜祖细胞池(RPC)产生的(Wallace,2011)。已经产生足够数量的特定细胞类型的RPC知道的一种方式是通过负反馈信号,其由分化细胞发射并且必须达到阈值水平以阻止该细胞类型的额外分化。评估负反馈信号是否由分化细胞发出的关键测定是异源沉淀测定,其中早期RPC被解离并用BrdU标记,然后与20倍过量的解离的分化细胞混合。然后将组合的细胞再次聚集并在细胞膜上培养7-10天。在这段时间内,RPC将会分化,BrdU + RPC的命运可以使用细胞类型特异性标记进行评估。开发这种沉淀测定的研究人员最初表明,当两种细胞类型混合在一起时,新生儿RPC与胚胎RPC相比,在加速进度条件下产生杆(Watanabe和Raff,1990; Watanabe等,1997)。我们已经使用这种测定来证明我们发现作为视网膜神经节细胞(RGC)分化的负调节物的声刺猬(Shh)促进RPC增殖(Jensen和Wallace,1997; Ringuette等,2014)。最近我们修改了异质性沉淀测定法,以评估视网膜无长突细胞的反馈信号的作用,将转化生长因子β2(Tgfβ2)鉴定为负反馈信号,并将Pten作为Tgfβ2应答的调节剂(Ma et al。,2007 ; ...

Ex vivo Culture of Adult Mouse Antral Glands
Author:
Date:
2017-01-05
[Abstract]  The tri-dimensional culture, initially described by Sato et al. (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker et al. (2010), who efficiently generated organoids that recapitulate the mature pyloric epithelium in vitro. This ex vivo approach is suitable and promising to study gastric function in homeostasis as well as in disease. We have adapted Barker’s protocol to compare homeostatic and regenerating tissues and here, we meticulously describe, step by step, the isolation and culture of antral glands as well as the isolation of single cells from antral glands that ... [摘要]  为了分离和表征成年小肠的上皮干细胞,最初由Sato等人(2009)描述的三维培养物已经随后适应于许多不同的器官。其中一个例子是Barker等人(2010)分离和培养窦性干细胞,他们有效地产生了在体外重现成熟幽门上皮的组织细胞。这种“离体”方法是适合的,并且有希望研究体内平衡和疾病中的胃功能。我们已经调整了Barker的方案来比较稳态和再生组织,这里,我们一步一步地仔细地描述了窦腺的分离和培养,以及从细胞分选后可能用于培养的窦腺中分离单细胞一个例子(Fernandez Vallone等人,2016)。

背景来自腺体的小鼠成体干细胞可以在3D matrigel中离体生长,作为“迷你腺体”无限期(Barker等人,2010) 。与EGF,Noggin和R-spondin 1存在下生长的小鼠成年小肠的干细胞相比,胃干细胞需要进一步补充Fgf10,胃泌素,Wnt3a和更高浓度的R-螺旋菌素1(称为作为ENRFGW)获得生产性文化。直到最近,在干细胞消融后,在离体培养系统中成体再生窦腺是否生长,如果是这样,仍然是未知的。使用本方案,证明了内源性和再生的腺体在接种时不会类似地生长并且表现出不同的生长培养要求。

Ex vivo Culture of Fetal Mouse Gastric Epithelial Progenitors
Author:
Date:
2017-01-05
[Abstract]  Isolation and tridimensional culture of murine fetal progenitors from the digestive tract represents a new approach to study the nature and the biological characteristics of these epithelial cells that are present before the onset of the cytodifferentiation process during development. In 2013, Mustata et al. described the isolation of intestinal fetal progenitors growing as spheroids in the ex vivo culture system initially implemented by Sato et al. (2009) to grow adult intestinal stem cells. Noteworthy, fetal-derived spheroids have high self-renewal capacity making easy their indefinite maintenance in culture. Here, we report an adapted protocol for isolation and ex vivo culture and maintenance of fetal epithelial progenitors from distal pre-glandular ... [摘要]  来自消化道的鼠胎儿祖细胞的分离和三维培养代表了研究在发育过程中细胞分化过程开始前存在的这些上皮细胞的性质和生物学特征的新方法。在2013年,Mustata等人描述了在最初由佐藤等人实施的离体培养系统中分离作为球体生长的肠胎细胞祖细胞。 >(2009)增长成年肠干细胞。值得注意的是,胎儿衍生的球体具有较高的自我更新能力,使其在文化中的无限期维护变得容易。在这里,我们报告了用于分离和远离前胃腺胃生长为胃球体的胎儿上皮祖细胞的分离和离体培养和维持的修改方案(Fernandez Vallone等人, 2016)。

背景 来自腺体的小鼠成体干细胞可以在3D matrigel中离体生长,作为“迷你腺体”无限期(Barker等人,2010) 。与在EGF,Noggin和R-spondin 1存在下生长的小肠的干细胞相比,成年胃干细胞需要进一步补充Fgf10,胃泌素,Wnt3a和更高浓度的R-spondin 1以获得生产性 - 文化。相比之下,到目前为止,很少知道在发育期间排列前腺上皮细胞的胎儿细胞。到目前为止,它们的性质以及其离体的潜在生长特性未明确。基于以前的研究,确定胎儿小肠(Mustata等人,2013年)中存在的细胞,我们报告了作为球体的小鼠胎儿胃祖细胞的培养(Fernandez Vallone et al。 。,2016)。可以在2009年由佐藤等人先前报道的培养基中重复胃祖细胞以生长小肠成体干细胞,与成人型胃干细胞相反,它们不需要额外的生长因子补充(Fgf10,Wnt3a或胃泌素)。 ...

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