Impact of Oxygen Concentration on Metabolic Profile of Human Placenta-Derived Mesenchymal Stem Cells As Determined by Chemical Isotope Labeling LC–MS,Journal of Proteome Research

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Impact of Oxygen Concentration on Metabolic Profile of Human Placenta-Derived Mesenchymal Stem Cells As Determined by Chemical Isotope Labeling LC–MS
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2018-04-26 , DOI: 10.1021/acs.jproteome.7b00887
Dan Wang ₁ , Deying Chen ₁ , Jiong Yu ₁ , Jingqi Liu ₁ , Xiaowei Shi ₂ , Yanni Sun ₁ , Qiaoling Pan ₁ , Xian Luo ₃ , Jinfeng Yang ₁ , Yang Li ₄ , Hongcui Cao ₁ , Liang Li ₃ , Lanjuan Li ₁

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The placenta resides in a physiologically low oxygen microenvironment of the body. Hypoxia induces a wide range of stem cell cellular activities. Here, we report a workflow for exploring the role of physiological (hypoxic, 5% oxygen) and original cell culture (normoxic, 21% oxygen) oxygen concentrations in regulating the metabolic status of human placenta-derived mesenchymal stem cells (hPMSCs). The general biological characteristics of hPMSCs were assessed via a variety of approaches such as cell counts, flow cytometry and differentiation study. A sensitive ¹³C/¹²C-dansyl labeling liquid chromatography–mass spectrometry (LC–MS) method targeting the amine/phenol submetabolome was used for metabolic profiling of the cell and corresponding culture supernatant. Multivariate and univariate statistical analyses were used to analyze the metabolomics data. hPMSCs cultured in hypoxia display smaller size, higher proliferation, greater differentiation ability and no difference in immunophenotype. Overall, 2987 and 2860 peak pairs or metabolites were detected and quantified in hPMSCs and culture supernatant, respectively. Approximately 86.0% of cellular metabolites and 84.3% of culture supernatant peak pairs were identified using a dansyl standard library or matched to metabolite structures using accurate mass search against human metabolome libraries. The orthogonal partial least-squares discriminant analysis (OPLS-DA) showed a clear separation between the hypoxic group and the normoxic group. Ten metabolites from cells and six metabolites from culture supernatant were identified as potential biomarkers of hypoxia. This study demonstrated that chemical isotope labeling LC–MS can be used to reveal the role of oxygen in the regulation of hPMSC metabolism, whereby physiological oxygen concentrations may promote arginine and proline metabolism, pantothenate and coenzyme A (CoA) biosynthesis, and alanine, aspartate and glutamate metabolism.

中文翻译：

化学同位素标记LC-MS确定氧浓度对人胎盘间充质干细胞代谢谱的影响

胎盘位于人体生理上低氧的微环境中。缺氧诱导了广泛的干细胞细胞活性。在这里，我们报告了一个工作流程，用于探索生理（低氧，5％氧气）和原始细胞培养物中（氧正常，21％氧气）氧气浓度在调节人胎盘来源的间充质干细胞（hPMSC）的代谢状态中的作用。hPMSC的一般生物学特性通过多种方法进行评估，例如细胞计数，流式细胞术和分化研究。敏感的¹³ C / ¹²针对胺/苯酚亚代谢组的C-丹磺酰基标记液相色谱-质谱（LC-MS）方法用于细胞和相应培养上清液的代谢分析。多变量和单变量统计分析用于分析代谢组学数据。在缺氧条件下培养的hPMSCs体积更小，增殖能力更高，分化能力更高，免疫表型无差异。总体而言，在hPMSC和培养上清液中分别检测到2987和2860个峰对或代谢物并进行了定量。使用dansyl标准文库鉴定出约86.0％的细胞代谢物和84.3％的培养上清液峰对，或使用针对人类代谢物文库的精确质量搜索，将其与代谢物结构进行匹配。正交偏最小二乘判别分析（OPLS-DA）显示了低氧组和常氧组之间的明确分离。来自细胞的十种代谢物和来自培养上清液的六种代谢物被鉴定为缺氧的潜在生物标记。这项研究表明，化学同位素标记LC-MS可用于揭示氧气在hPMSC代谢调节中的作用，由此生理性氧气浓度可促进精氨酸和脯氨酸代谢，泛酸和辅酶A（CoA）的生物合成，以及丙氨酸，天冬氨酸和谷氨酸代谢。

更新日期：2018-04-26

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