Huang Lei, Niu Chenguang, Willard Belinda, Zhao Weimin, Liu Lan, He Wei, Wu Tianwen, Yang Shulin, Feng Shutang, Mu Yulian, Zheng Lemin, Li Kui
State Key Laboratory of Animal Nutrition and Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Haidian District 100193, Beijing, China.
The Institute of Cardiovascular Sciences and Institute of Systems Biomedicine, School of Basic Medical Sciences, and Key Laboratory of Molecular Cardiovascular Sciences of Ministry of Education, Peking University Health Science Center, No. 38 Xueyuan Road, Haidian District 100191, Beijing, China.
Stem Cell Res Ther. 2015 Apr 15;6(1):77. doi: 10.1186/s13287-015-0061-x.
Mesenchymal stem cells (MSCs) have the ability to proliferate in vivo with a large variety of differentiation potentials and therefore are widely used as an ideal material for cell therapy. MSCs derived from pig and human sources are similar in many aspects, such as cell immunophenotype and functional characteristics. However, differences in proteomics and the molecular mechanisms of cell functions between porcine bone marrow MSCs (BM-MSCs) and umbilical cord MSCs (UC-MSCs) are largely unknown. To the best of our knowledge, MSCs collected from different tissue have specific phenotype and differentiation ability in response to microenvironment, known as a niche.
Porcine BM-MSCs and UC-MSCs were evaluated with flow cytometric and adipogenic and osteogenic differentiation analyses. We used isobaric tagging for relative and absolute quantitation (iTRAQ), combined with liquid chromatography-tandem mass spectrometry, to identify differentially expressed proteins (DEPs) between these two types of MSCs. Kyoto Encyclopedia of Genes and Genomes pathway and phenotype analyses were used to understand the links between cell migration ability and DEPs.
Two separate iTRAQ experiments were conducted, identifying 95 DEPs (95% confidence interval). Five of these proteins were verified by Western blotting. These 95 DEPs were classified in terms of biological regulation, metabolic process, developmental process, immune system process, reproduction, death, growth, signaling, localization, response to stimulus, biological adhesion, and cellular component organization. Our study is the first to show results indicating that porcine BM-MSCs have a higher migration capability than UC-MSCs. Finally, one of the DEPs, Vimentin, was verified to have a positive role in MSC migration.
These results represent the first attempt to use proteomics specifically targeted to porcine MSCs of different tissues. The identified components should help reveal a variety of tissue-specific functions in tissue-derived MSC populations and could serve as important tools for the regeneration of particular tissues in future stem cell-based tissue engineering studies using animal models.
间充质干细胞（MSCs）具有在体内增殖的能力，并具有多种分化潜能，因此被广泛用作细胞治疗的理想材料。猪源和人源的间充质干细胞在许多方面相似，如细胞免疫表型和功能特性。然而，猪骨髓间充质干细胞（BM-MSCs）和脐带间充质干细胞（UC-MSCs）之间蛋白质组学和细胞功能分子机制的差异在很大程度上尚不清楚。据我们所知，从不同组织收集的间充质干细胞在响应微环境（即生态位）时具有特定的表型和分化能力。
采用流式细胞术、脂肪生成和成骨分化分析对猪BM-MSCs和UC-MSCs进行评估。我们使用相对和绝对定量的等压标记（iTRAQ）结合液相色谱-串联质谱来鉴定这两种间充质干细胞之间的差异表达蛋白（DEPs）。利用京都基因与基因组百科全书通路和表型分析来了解细胞迁移能力与DEPs之间的联系。
进行了两项独立的iTRAQ实验，鉴定出95个DEPs（95%置信区间）。其中5种蛋白质通过蛋白质免疫印迹法得到验证。这95个DEPs根据生物调节、代谢过程、发育过程、免疫系统过程、繁殖、死亡、生长、信号传导、定位、对刺激的反应、生物粘附和细胞成分组织进行分类。我们的研究首次表明猪BM-MSCs比UC-MSCs具有更高的迁移能力。最后，其中一个DEPs波形蛋白被证实对间充质干细胞迁移具有积极作用。
这些结果代表了首次针对不同组织的猪间充质干细胞进行蛋白质组学研究的尝试。所鉴定的成分应有助于揭示组织来源的间充质干细胞群体中各种组织特异性功能，并可作为未来基于动物模型的干细胞组织工程研究中特定组织再生的重要工具。
