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Effect of Different Preconditioning Regimens on the Expression Profile of Murine Adipose-Derived Stromal/Stem Cells

机译:不同预处理方案对小鼠脂肪基质/干细胞表达谱的影响

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摘要

Stem cell-based therapies require cells with a maximum regenerative capacity in order to support regeneration after tissue injury and organ failure. Optimization of this regenerative potential of mesenchymal stromal/stem cells (MSC) or their conditioned medium by in vitro preconditioning regimens are considered to be a promising strategy to improve the release of regenerative factors. In the present study, MSC were isolated from inguinal adipose tissue (mASC) from C57BL/6 mice, cultured, and characterized. Then, mASC were either preconditioned by incubation in a hypoxic environment (0.5% O2), or in normoxia in the presence of murine epidermal growth factor (EGF) or tumor necrosis factor α (TNFα) for 48 h. Protein expression was measured by a commercially available array. Selected factors were verified by PCR analysis. The expression of 83 out of 308 proteins (26.9%) assayed was found to be increased after preconditioning with TNFα, whereas the expression of 61 (19.8%) and 70 (22.7%) proteins was increased after incubation with EGF or in hypoxia, respectively. Furthermore, we showed the proliferation-promoting effects of the preconditioned culture supernatants on injured epithelial cells in vitro. Our findings indicate that each preconditioning regimen tested induced an individual expression profile with a wide variety of factors, including several growth factors and cytokines, and therefore may enhance the regenerative potential of mASC for cell-based therapies.
机译:基于干细胞的疗法需要细胞具有最大的再生能力,以支持组织损伤和器官衰竭后的再生。通过体外预处理方案优化间充质基质/干细胞(MSC)或它们的条件培养基的这种再生潜能被认为是改善再生因子释放的一种有前途的策略。在本研究中,从C57BL / 6小鼠的腹股沟脂肪组织(mASC)中分离出MSC,并对其进行了培养和鉴定。然后,通过在缺氧环境(0.5%O2)中孵育或在鼠类表皮生长因子(EGF)或肿瘤坏死因子α(TNFα)存在的常氧条件下对mASC进行预处理48小时。通过可商购的阵列测量蛋白质表达。通过PCR分析验证选择的因素。用TNFα预处理后,发现308种蛋白质中83种的蛋白表达(26.9%)增加,而EGF孵育或缺氧后分别增加61种蛋白(19.8%)和70种蛋白(22.7%)的表达。 。此外,我们显示了预处理培养物上清液对体外损伤的上皮细胞的增殖促进作用。我们的发现表明,所测试的每种预处理方案均诱导了具有多种因素的个体表达谱,其中包括多种生长因子和细胞因子,因此可能增强mASC在基于细胞的治疗中的再生潜力。

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