GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells
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Author:
Date:
2017-12-05
[Abstract] Human induced pluripotent stem cells (hiPSCs) are a promising tool in cell-based therapies for degenerative diseases. A safe application of hiPSCs in vivo, requires the detection of the presence of residual undifferentiated pluripotent cells that can potentially cause the insurgence of teratomas. Several studies point out that metabolic products may provide an alternative method to identify the different steps of cells differentiation. In particular, the analysis of volatile organic compounds (VOCs) is gaining a growing interest in this context, thanks to its inherent noninvasiveness. Here, a protocol for VOCs analysis from human induced pluripotent stem cells (hiPSCs) is illustrated. It is based on Solid-Phase Microextraction (SPME) technique coupled with gas chromatography-mass ...
[摘要] 人诱导的多能干细胞(hiPSC)是用于退化性疾病的基于细胞的疗法中的有前景的工具。 hiPSCs在体内的安全应用需要检测残留未分化多能细胞的存在,这可能会导致畸胎瘤的爆发。几项研究指出,代谢产物可能提供了另一种方法来确定细胞分化的不同步骤。特别是挥发性有机化合物(VOCs)的分析由于其固有的非侵入性而在这方面越来越受到关注。在这里,说明了从人诱导的多能干细胞(hiPSC)分析VOC的方案。它基于固相微萃取(SPME)技术与气相色谱 - 质谱联用(GC / MS)。该方法用于测量从绒毛膜样品(CVS)到hiPSC的细胞重编程期间和沿着hiPSC体外分化成早期神经祖细胞(NP)的细胞顶空中的挥发性代谢物修饰,穿过胚状体机构(EBs)的形成。
【背景】提出细胞代谢作为在分化的各个步骤期间研究干细胞的替代物。事实上,假设干细胞从多能性向完全分化的转变可能引起代谢产物的剧烈变化是合理的。在诱导的多能干细胞,亲本成纤维细胞和胚胎干细胞之间观察到了这种假设的第一个证据(Meissen等人,2012)。
在代谢产物中,挥发性有机化合物(VOC)吸引了人们对其收集的简单性,内在的非侵入性和广泛的分析方法的广泛关注(Boots et。,2015 ...
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Isolation of Intestinal Mesenchymal Cells from Adult Mice
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Author:
Date:
2016-09-20
[Abstract] During the last 20 years intestinal mesenchymal cells (IMCs) have emerged as an important cell type that plays a central role in intestinal development and homeostasis, by providing both structural support and growth regulatory elements. IMCs also actively participate in wound healing responses, thus regulating pathologic conditions such as tissue repair, inflammation, fibrosis and carcinogenesis (Powell et al., 2011). We have recently demonstrated that intestinal mesenchymal-specific signals play important in vivo physiological roles in intestinal inflammation and carcinogenesis (Koliaraki et al., 2012; Roulis et al., 2014; Koliaraki et al., 2015). Here we describe the enzymatic method used for the isolation and culture of mesenchymal cells ...
[摘要] 在过去20年间,肠间质细胞(IMC)已经作为重要的细胞类型出现,通过提供结构支持和生长调节元件在肠发育和体内平衡中起着中心作用。 IMC还积极参与伤口愈合反应,从而调节病理状况,例如组织修复,炎症,纤维化和癌发生(Powell等人,2011)。 我们最近已经证明肠间充质特异性信号在肠炎症和癌发生中在体内起重要的生理作用(Koliaraki等人,2012; Roulis等人,/em>。,2014; Koliaraki 。。,2015)。 在这里我们描述了用于从成年小鼠肠道分离和培养间充质细胞的酶法。
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Preparation of Single Cell Suspensions from Mouse Aorta
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Author:
Date:
2016-06-05
[Abstract] Atherosclerosis is a chronic inflammatory disease of the arterial wall characterized by lipid deposition, plaque formation, and immune cell infiltration. Innate and adaptive immune cells infiltrate the artery during development of the disease. Moreover, advanced disease leads to formation of artery tertiary lymphoid organs in the adventitia (Grabner et al., 2009; Hu et al., 2015). Various and diverse types of immune cells have been identified in the aorta adventitia vs atherosclerotic plaques (Elewa et al., 2016; Galkina et al., 2006; Lotzer et al., 2010; Mohanta et al., 2016; Mohanta et al., 2014; Moos et al., 2005; Srikakulapu et al., 2016; Zhao et al., 2004). There are conflicting reports on the number and subtypes ...
[摘要] 动脉粥样硬化是动脉壁的慢性炎性疾病,其特征在于脂质沉积,斑块形成和免疫细胞浸润。先天性和适应性免疫细胞在疾病发展期间浸润动脉。此外,晚期疾病导致外膜中动脉三级淋巴器官的形成(Grabner等人,2009; Hu等人,2015)。已经在主动脉外膜vs动脉粥样硬化斑块中鉴定了各种不同类型的免疫细胞(Elewa等人,2016; Galkina等人,2006; Lotzer等人, 2010; Mohanta等人,2016; Mohanta等人,2014; Moos等人,2010; Mohanta等人,2010; 2005; Srikakulapu et al。,2016; Zhao et al。,2004)。根据动物的年龄,用于获得单细胞悬浮液的方案和小鼠的饮食条件,存在关于主动脉中免疫细胞的数量和亚型的矛盾报告(Campbell等, 2012; Clement等人,2015; Galkina等人,2006; Kyaw等人,2012)。使用不同的方案,主动脉中免疫细胞的数目差异多达十倍(Butcher等,2012; Galkina等,2006; Gjurich等,2015; Grabner等人,2009; ...
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