A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells
A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells
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使用小鼠脂肪来源的间充质干细胞对原代培养细胞和传代培养细胞进行基于串联质谱的液相色谱蛋白质组分析
DOI：
10.1155/2019/7274057
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发表时间：
2019
影响因子：
4.3
通讯作者：
Noguchi Hirofumi
中科院分区：
文献类型：
--
作者：
Nakashima Yoshiki;Nahar Saifun;Miyagi-Shiohira Chika;Kinjo Takao;Kobayashi Naoya;Saitoh Issei;Watanabe Masami;Fujita Jiro;Noguchi Hirofumi
脂肪间充质干细胞(MSC-ATS)是细胞治疗的代表性细胞来源。然而，传代引起的细胞应激如何影响MSC-AT蛋白的表达还不清楚。本研究以小鼠原代培养细胞(P0)和传代3次的细胞(P3)为研究对象，采用LC-MS/MS联用技术进行蛋白质表达分析。在P0中，共有256个蛋白质被归类为细胞过程相关蛋白，179个被归类为代谢过程相关蛋白。这些被认为是细胞对体外环境的适应性反应。然而，在P0中发现了7个与生长有关的蛋白(CSF1、App、Adam15、Alcam、Tbl1xr1、Nins1和SBDS)。此外，在P0中还鉴定了四种具有抗氧化活性的蛋白质(Srxn1、Txndc17、Fam213b和APOE)。我们鉴定了在P0和P3细胞中表达的1139个蛋白质，其中P3细胞的表达比P0细胞减少了69.4%，但1139个蛋白质很可能是来源于MSC-AT的蛋白质。MSC-ATS在传代3代后仍保持功能。然而，LC-MS/MS分析数据表明，经过三代后，蛋白质的表达有所下降。MSC-ATS在P3细胞中保持了约70%的蛋白表达能力。
Adipose‐derived mesenchymal stem cells (MSC‐ATs) are representative cell sources for cell therapy. However, how cell stress resulting from passage influences the MSC‐AT protein expression has been unclear. In this study, a protein expression analysis was performed by liquid chromatography with tandem mass spectrometry (LC‐MS/MS) using mouse primary cultured cells (P0) and cells passaged three times (P3) as samples. A total of 256 proteins were classified as cellular process‐related proteins, while 179 were classified as metabolic process‐related proteins in P0. These were considered to be adaptive responses of the cells to an in vitro environment. However, seven proteins of growth were identified (Csf1, App, Adam15, Alcam, Tbl1xr1, Ninj1, and Sbds) in P0. In addition, four proteins of antioxidant activity were also identified (Srxn1, Txndc17, Fam213b, and Apoe) in P0. We identified 1139 proteins expressed in both P0 and P3 cells that had their expression decreased to 69.4% in P3 cells compared with P0 cells, but 1139 proteins are very likely proteins that are derived from MSC‐AT. The function of MSC‐ATs was maintained after three passages. However, the LC‐MS/MS analysis data showed that the protein expression was degraded after three passages. MSC‐ATs retained about 70% of their protein expression ability in P3 cells.