A liquid chromatography with tandem mass spectrometry-based proteomic analysis of primary cultured cells and subcultured cells using mouse adipose-derived mesenchymal stem cells

Yoshiki Nakashima, Saifun Nahar, Chika Miyagi-Shiohira, Takao Kinjo, Naoya Kobayashi, Issei Saitoh, Masami Watanabe, Jiro Fujita, Hirofumi Noguchi

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Article number7274057
JournalStem Cells International
Volume2019
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Tandem Mass Spectrometry
Mesenchymal Stromal Cells
Liquid Chromatography
Proteomics
Cultured Cells
Proteins
Myelin P0 Protein
Apolipoproteins E
Cell- and Tissue-Based Therapy
Antioxidants

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

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.

In: Stem Cells International, Vol. 2019, 7274057, 01.01.2019.

Research output: Contribution to journalArticle

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. In: Stem Cells International. 2019 ; Vol. 2019.
@article{d0baae753b5f492cbdf478a3c3295b65,
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",
abstract = "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.",
author = "Yoshiki Nakashima and Saifun Nahar and Chika Miyagi-Shiohira and Takao Kinjo and Naoya Kobayashi and Issei Saitoh and Masami Watanabe and Jiro Fujita and Hirofumi Noguchi",
year = "2019",
month = "1",
day = "1",
doi = "10.1155/2019/7274057",
language = "English",
volume = "2019",
journal = "Stem Cells International",
issn = "1687-9678",
publisher = "Hindawi Publishing Corporation",

}

TY - JOUR

T1 - A liquid chromatography with tandem mass spectrometry-based proteomic analysis of primary cultured cells and subcultured cells using mouse adipose-derived mesenchymal stem cells

AU - Nakashima, Yoshiki

AU - Nahar, Saifun

AU - Miyagi-Shiohira, Chika

AU - Kinjo, Takao

AU - Kobayashi, Naoya

AU - Saitoh, Issei

AU - Watanabe, Masami

AU - Fujita, Jiro

AU - Noguchi, Hirofumi

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85065823010&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85065823010&partnerID=8YFLogxK

U2 - 10.1155/2019/7274057

DO - 10.1155/2019/7274057

M3 - Article

AN - SCOPUS:85065823010

VL - 2019

JO - Stem Cells International

JF - Stem Cells International

SN - 1687-9678

M1 - 7274057

ER -