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Gene expression and protein secretion during human mesenchymal cell differentiation into adipogenic cells

机译:人间充质细胞向成脂细胞分化过程中的基因表达和蛋白分泌

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Background Mesenchymal stromal cells (MSC) can be obtained from potentially any tissue from the human body, but cells purified from different sources are undoubtedly different, and for each medical application, the MSC with the best regenerative potential should be chosen. Results Bone marrow-derived mesenchymal stromal cells (BM-MSC), adipose tissue-derived mesenchymal stromal cells (AT-MSC) and Wharton’s Jelly-derived mesenchymal stromal cells (WJ-MSC) were isolated from human tissues and were cultured under differentiation media supplemented with fetal bovine serum. We quantified the expression of stem cell and adipocyte genetic markers using quantitative real time PCR, as well as the secretion of cytokines, extracellular matrix components and growth factors using Luminex and ELISA. All three MSC differentiated into adipogenic cells. AT-MSC showed the highest shift in ADIPOQ, CEBPA and PPARG mRNA expression. BM-MSC kept high expression levels of stem-cell markers SOX2 and POU5F1. WJ-MSC showed the lowest increase in mRNA expression when cells were induced to differentiate into adipocytes. Regarding protein secretion, adipocyte-like cells generated from WJ-MSC secreted the highest chemokine levels. AT-MSC-derived adipocyte-like cells secreted the lowest cytokine amounts and the highest quantity of collagen types I and III. Adipocyte-like cells obtained from BM-MSC secreted high amounts of most angiogenic factors, growth factors TGF-β1 and TGF-β2, collagens type II and IV, heparan sulfate, laminin and aggrecan. Conclusion Mesenchymal stromal cells purified from different tissues have a different behavior when induced to differentiate into adipocyte-like cells.
机译:背景间充质基质细胞(MSC)可以从人体的任何组织中获得,但从不同来源纯化的细胞无疑是不同的,对于每种医学应用,应选择具有最佳再生潜能的MSC。结果从人组织中分离得到骨髓来源的间充质基质细胞(BM-MSC),脂肪组织来源的间质基质细胞(AT-MSC)和沃顿氏胶质来源的间质基质细胞(WJ-MSC),并在分化培养基下培养。补充胎牛血清。我们使用定量实时PCR定量了干细胞和脂肪细胞遗传标志物的表达,并使用Luminex和ELISA定量了细胞因子,细胞外基质成分和生长因子的分泌。所有三个MSC均分化为成脂细胞。 AT-MSC在ADIPOQ,CEBPA和PPARG mRNA表达中表现出最高的变化。 BM-MSC保持了干细胞标记SOX2和POU5F1的高表达水平。当诱导细胞分化为脂肪细胞时,WJ-MSC显示出最低的mRNA表达增加。关于蛋白质分泌,由WJ-MSC产生的脂肪细胞样细胞分泌最高的趋化因子水平。 AT-MSC衍生的脂肪细胞样细胞分泌的细胞因子含量最低,而I型和III型胶原蛋白的分泌最高。从BM-MSC获得的脂肪细胞样细胞分泌大量的大多数血管生成因子,生长因子TGF-β1和TGF-β2,II型和IV型胶原,硫酸乙酰肝素,层粘连蛋白和聚集蛋白聚糖。结论从不同组织中纯化的间充质基质细胞在诱导分化为脂肪细胞样细胞时具有不同的行为。

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