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人骨髓间充质细胞向脂肪生成细胞分化过程中的基因表达与蛋白质分泌

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

作者信息

Amable Paola Romina, Teixeira Marcus Vinicius Telles, Carias Rosana Bizon Vieira, Granjeiro José Mauro, Borojevic Radovan

机构信息

Excellion Biomedical Services S.A., Rua Afrânio de Mello Franco 333, Quitandinha, Petrópolis, Rio de Janeiro, Brazil.

National Institute of Metrology, Quality and Technology (Inmetro), Xerém, Rio de Janeiro, Brazil.

出版信息

BMC Cell Biol. 2014 Dec 20;15:46. doi: 10.1186/s12860-014-0046-0.

Abstract

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.

摘要

背景

间充质基质细胞(MSC)几乎可从人体的任何组织中获取,但从不同来源纯化得到的细胞无疑存在差异,对于每种医学应用,都应选择具有最佳再生潜能的MSC。

结果

从人体组织中分离出骨髓来源的间充质基质细胞(BM-MSC)、脂肪组织来源的间充质基质细胞(AT-MSC)和脐带来源的间充质基质细胞(WJ-MSC),并在添加胎牛血清的分化培养基中进行培养。我们使用定量实时PCR对干细胞和脂肪细胞遗传标志物的表达进行定量,同时使用Luminex和ELISA对细胞因子、细胞外基质成分和生长因子的分泌进行定量。所有三种MSC均分化为脂肪生成细胞。AT-MSC在ADIPOQ、CEBPA和PPARG mRNA表达方面表现出最大变化。BM-MSC保持干细胞标志物SOX2和POU5F1的高表达水平。当细胞被诱导分化为脂肪细胞时,WJ-MSC的mRNA表达增加最少。在蛋白质分泌方面,WJ-MSC产生的脂肪细胞样细胞分泌的趋化因子水平最高。AT-MSC来源的脂肪细胞样细胞分泌的细胞因子量最低,I型和III型胶原蛋白量最高。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型胶原蛋白、硫酸乙酰肝素、层粘连蛋白和聚集蛋白聚糖。

结论

从不同组织纯化得到的间充质基质细胞在被诱导分化为脂肪细胞样细胞时具有不同的行为。

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