Maia Leandro, de Moraes Carolina Nogueira, Dias Marianne Camargos, Martinez Julia Bauzá, Caballol Antonia Odena, Testoni Giorgia, de Queiroz Carla Martins, Peña Ramón Díaz, Landim-Alvarenga Fernanda C, de Oliveira Eliandre
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, São Paulo State University UNESP, Botucatu, São Paulo 18618-681, Brazil; Proteomics Platform, Parc Cientific de Barcelona (PCB), Barcelona 08028, Spain.
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, São Paulo State University UNESP, Botucatu, São Paulo 18618-681, Brazil.
Theriogenology. 2017 Sep 15;100:8-15. doi: 10.1016/j.theriogenology.2017.05.015. Epub 2017 May 24.
To the best of our knowledge, this is the first study describing the proteome of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs) in a global and functional manner. The aim of this work was to analyze the proteome of previously characterized UCIM-MSCs to determine protein abundance and classify the identified proteins according to Gene Ontology (GO) terms. Protein classification analysis according to biological process, molecular function and cellular component was performed using the PANTHER (Protein ANalysis THrough Evolutionary Relationships) Classification System, which revealed enrichment for 42 biological processes, 23 molecular functions and 18 cellular components. Protein abundance was estimated according to the emPAI method (Exponential Modified Protein Abundance Index). The two most abundant proteins in the proteome of UCIM-MSCs were the cytoskeletal proteins actin and vimentin, which have important roles in cell stability and motility. Additionally, we identified 14 cell surface antigens. Three of them, CD44, CD90 and CD105, had been previously validated by flow cytometry. In the present study, we also identified important information about the biological properties of UCIM-MSCs such as differentiation potential, low immunogenicity (low MHC-II expression) and chromosomal stability, which reinforces their use for cell therapy. Together with the proteomic findings, this information allowed us to infer the functional relevance of several activities related to primary metabolic processes, protein synthesis, production of vesicle coats, vesicle-mediated transport and antioxidant activity. In addition, the identification of different cell surface markers may help establish an immunophenotypic panel suitable for the characterization of MSCs from equine fetal membranes.
据我们所知,这是第一项以全面且功能化的方式描述马脐带血管间基质间充质干细胞(UCIM-MSCs)蛋白质组的研究。这项工作的目的是分析先前已鉴定的UCIM-MSCs的蛋白质组,以确定蛋白质丰度,并根据基因本体论(GO)术语对鉴定出的蛋白质进行分类。使用PANTHER(通过进化关系进行蛋白质分析)分类系统对蛋白质进行了基于生物学过程、分子功能和细胞成分的分类分析,结果显示42个生物学过程、23个分子功能和18个细胞成分显著富集。根据emPAI方法(指数修正蛋白质丰度指数)估算蛋白质丰度。UCIM-MSCs蛋白质组中含量最高的两种蛋白质是细胞骨架蛋白肌动蛋白和波形蛋白,它们在细胞稳定性和运动性中发挥着重要作用。此外,我们鉴定出了14种细胞表面抗原。其中三种,即CD44、CD90和CD105,先前已通过流式细胞术验证。在本研究中,我们还确定了关于UCIM-MSCs生物学特性的重要信息,如分化潜能、低免疫原性(低MHC-II表达)和染色体稳定性,这进一步证明了它们在细胞治疗中的应用价值。结合蛋白质组学研究结果,这些信息使我们能够推断出与初级代谢过程、蛋白质合成、囊泡衣的产生、囊泡介导的运输和抗氧化活性相关的几种活动的功能相关性。此外,不同细胞表面标志物的鉴定可能有助于建立一个适合表征马胎膜间充质干细胞的免疫表型组。