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Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham

Company: Sigma-Aldrich
Catalog#: D6421
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GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells
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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 ...

Teratoma Formation Assay for Assessing Pluripotency and Tumorigenicity of Pluripotent Stem Cells
Author:
Date:
2017-08-20
[Abstract]  Pluripotent stem cells such as induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) form teratomas when transplanted into immunodeficient mice. As teratomas contain all three germ layers (endoderm, mesoderm, ectoderm), teratoma formation assay is widely used as an index of pluripotency (Evans and Kaufman, 1981; Hentze et al., 2009; Gropp et al., 2012). On the other hand, teratoma-forming tumorigenicity also represents a major risk factor impeding potential clinical applications of pluripotent stem cells (Miura et al., 2009; Okano et al., 2013). Recently, we reported that iPSCs derived from naked mole-rat lack teratoma-forming tumorigenicity when engrafted into the testes of non-obese diabetic/severe combined immunodeficient (NOD/SCID) ... [摘要]  多能干细胞,如诱导多能干细胞(iPSCs)和胚胎干细胞(ESC),当移植到免疫缺陷小鼠时,形成畸胎瘤。由于畸胎瘤包含所有三个胚层(内胚层,中胚层,外胚层),畸胎瘤形成测定被广泛用作多能性的指标(Evans和Kaufman,1981; Hentze等,2009; Gropp等,2012)。另一方面,畸胎瘤形成致瘤性也是阻碍多能干细胞潜在临床应用的主要危险因素(Miura et al。,2009; Okano等,2013)。最近,我们报道了由于ES细胞表达的Ras(ERAS)和替代物,嫁接到非肥胖型糖尿病/严重联合免疫缺陷型(NOD / SCID)小鼠的睾丸中,从裸鼠睾丸衍生的iPSC缺乏畸胎瘤形成致瘤性阅读框(ARF)依赖于该物种特异性的肿瘤抑制机制(Miyawaki等,2016)。在这里,我们描述了将多能干细胞移植到NOD / SCID小鼠的睾丸中以产生用于评估多能性和致瘤性的畸胎瘤的方法。
【背景】iPSCs和ESC用于再生医学细胞移植治疗中的应用。然而,当移植到免疫缺陷小鼠中时,这些细胞形成称为含有分化组织的畸胎瘤的肿瘤。因此,其畸胎瘤形成致瘤性的风险限制了其临床应用。几项研究报道了克服畸胎瘤形成肿瘤发生风险的方法(Itakura et al。,2017; Vazquez-Martin et ...

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