A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells
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 tande...
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/7274057 |
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| author | Yoshiki Nakashima Saifun Nahar Chika Miyagi-Shiohira Takao Kinjo Naoya Kobayashi Issei Saitoh Masami Watanabe Jiro Fujita Hirofumi Noguchi |
| author_facet | Yoshiki Nakashima Saifun Nahar Chika Miyagi-Shiohira Takao Kinjo Naoya Kobayashi Issei Saitoh Masami Watanabe Jiro Fujita Hirofumi Noguchi |
| author_sort | Yoshiki Nakashima |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-d215bb026a3b4e7f8f01f6d49b173386 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-d215bb026a3b4e7f8f01f6d49b1733862025-02-03T01:22:03ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/72740577274057A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem CellsYoshiki Nakashima0Saifun Nahar1Chika Miyagi-Shiohira2Takao Kinjo3Naoya Kobayashi4Issei Saitoh5Masami Watanabe6Jiro Fujita7Hirofumi Noguchi8Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, JapanDepartment of Infectious, Respiratory, and Digestive Medicine, University of the Ryukyus, Okinawa 903-0215, JapanDepartment of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, JapanDepartment of Basic Laboratory Sciences, School of Health Sciences in Faculty of Medicine, University of the Ryukyus, Okinawa 903-0215, JapanOkayama Saidaiji Hospital, Okayama 704-8192, JapanDivision of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, Niigata 951-8514, JapanDepartment of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, JapanDepartment of Infectious, Respiratory, and Digestive Medicine, University of the Ryukyus, Okinawa 903-0215, JapanDepartment of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Okinawa 903-0215, JapanAdipose-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.http://dx.doi.org/10.1155/2019/7274057 |
| spellingShingle | Yoshiki Nakashima Saifun Nahar Chika Miyagi-Shiohira Takao Kinjo Naoya Kobayashi Issei Saitoh Masami Watanabe Jiro Fujita Hirofumi Noguchi A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells Stem Cells International |
| title | A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells |
| title_full | A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells |
| title_fullStr | A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells |
| title_full_unstemmed | A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells |
| title_short | A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells |
| title_sort | liquid chromatography with tandem mass spectrometry based proteomic analysis of primary cultured cells and subcultured cells using mouse adipose derived mesenchymal stem cells |
| url | http://dx.doi.org/10.1155/2019/7274057 |
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