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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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Autores principales: Nakashima, Yoshiki, Nahar, Saifun, Miyagi-Shiohira, Chika, Kinjo, Takao, Kobayashi, Naoya, Saitoh, Issei, Watanabe, Masami, Fujita, Jiro, Noguchi, Hirofumi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362508/
https://www.ncbi.nlm.nih.gov/pubmed/30805011
http://dx.doi.org/10.1155/2019/7274057
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author Nakashima, Yoshiki
Nahar, Saifun
Miyagi-Shiohira, Chika
Kinjo, Takao
Kobayashi, Naoya
Saitoh, Issei
Watanabe, Masami
Fujita, Jiro
Noguchi, Hirofumi
author_facet Nakashima, Yoshiki
Nahar, Saifun
Miyagi-Shiohira, Chika
Kinjo, Takao
Kobayashi, Naoya
Saitoh, Issei
Watanabe, Masami
Fujita, Jiro
Noguchi, Hirofumi
author_sort Nakashima, Yoshiki
collection PubMed
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.
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spelling pubmed-63625082019-02-25 A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Primary Cultured Cells and Subcultured Cells Using Mouse Adipose-Derived Mesenchymal Stem Cells Nakashima, Yoshiki Nahar, Saifun Miyagi-Shiohira, Chika Kinjo, Takao Kobayashi, Naoya Saitoh, Issei Watanabe, Masami Fujita, Jiro Noguchi, Hirofumi Stem Cells Int Research Article 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. Hindawi 2019-01-10 /pmc/articles/PMC6362508/ /pubmed/30805011 http://dx.doi.org/10.1155/2019/7274057 Text en Copyright © 2019 Yoshiki Nakashima et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Nakashima, Yoshiki
Nahar, Saifun
Miyagi-Shiohira, Chika
Kinjo, Takao
Kobayashi, Naoya
Saitoh, Issei
Watanabe, Masami
Fujita, Jiro
Noguchi, Hirofumi
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 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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6362508/
https://www.ncbi.nlm.nih.gov/pubmed/30805011
http://dx.doi.org/10.1155/2019/7274057
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