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The Biomarkers and Variation with Adipogenic Differentiation of Mesenchymal Stem Cells Studied by in Vitro 9.4T MR Spectroscopy

Author: XuZhiFeng
Tutor: WuRenHua
School: Shantou University
Course: Medical Imaging and Nuclear Medicine
Keywords: Magnetic Resonance Spectroscopy Mesenchymal stem cells Adipogenic differentiation Biology logo
CLC: R329
Type: Master's thesis
Year: 2010
Downloads: 27
Quote: 0
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Abstract


Objective: 9.4T from Body high-resolution nuclear magnetic resonance spectrum to explore the human derived from umbilical cord the Waldorf-through plastic mesenchymal stem cell metabolism biomarkers characteristic simultaneous quantitative analysis of mesenchymal stem cells induced into adipogenic differentiation metabolite changes . METHODS: Cultured Collect 4th generation fusion rate of about 80-85% of mesenchymal stem cells and human esophageal cancer EC109 cells (approximately 7-8 × 106, the number of samples were n = 6 and n = 2; primary source of cells were multidisciplinary Center of Shantou University and Shantou University Medical College pathological laboratory). The final concentration of 1μM dexamethasone, 5001μM isobutyl xanthine, 60μM indole indomethacin and 5μg/mL of insulin in 10% FBS / LG-DMEM medium and 14-Tiancheng fat differentiation induced (sample of mesenchymal stem cells number n = 5); using Oil Red O staining and RT-PCR validation adipogenic differentiation model of success. 1H-MRS data acquisition from Body 9.4T high resolution magnetic resonance spectrum analyzer (Bruker Avance 400MHz). The acquired spectral data in the frequency domain by XWINNMR (Bruker GmBH) after the initial processing of the software, then Application Mestre-c 4.7 software for analysis. Results: mesenchymal stem cells obtained spectral quality and reproducibility. Mesenchymal stem cells in the spectrum visible main metabolite comprising: choline, creatine, glutamic acid, inositol, acetic acid, threonine, methionine, and fatty acid (1.28ppmbiomarker are considered to be neural stem cell specific ID). Before and after differentiation of mesenchymal stem cell adipogenic visible choline, creatine, glutamic acid, acetic acid metabolite were decreased, threonine, methionine and fatty acids actually was an upward trend. Quantitative analysis of total choline, acetic acid, glutamic acid and creatine, respectively, decreased from 6.3 ± 0.68,0.97 ± 0.23,0.3 ± 0.05and 0.1 ± 0.02 mM to 1.1 ± 0.06 (p lt; 0.01), 0.45 ± 0.1 (p lt; 0.01), 0.16 ± 0.08mM (p lt; 0.05) and could not be detected level; methionine, threonine and fatty acid concentrations of 1.28 ppm from 0.03 ± 0.01,0.11 ± 0.02 and (0.66 ± 0.17) n mM increased to 0.12 ± 0.05 (p lt; 0.01) and 0.15 ± 0.05 (p gt; 0.05) and (1.18 ± 0.07) n mM (p lt; 0.01) (n behalf of the methylene number). Adipogenic differentiation 14 days after the visible cell rounding, package quality of a lot of fat droplets formed Oil Red O staining fat droplets orange-red; reverse transcriptase PCR (RT-PCR), peroxisome proliferator-activated by visible body-r (PPAR-r), acyl-coenzyme A synthetase (ACS) and lipoprotein lipase (LPL) gene expression was significantly increased. Conclusion: The high-resolution magnetic resonance spectrum from the Body can obtain mesenchymal stem cell metabolic characteristics; mesenchymal stem cells also contain 1.28 ppm biomarker specific identification of neural stem cells, but it is not a stem cell-specific biological identity. According to the characteristics of changes in metabolites, magnetic resonance spectrum between recharge monitoring and identification of adipogenic differentiation of stem cells.

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