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.
Highlights
Mesenchymal stromal stem cells (MSCs) are considered to have the ability to differentiate into mesenchymal cells, such as osteoblasts, adipocytes, muscle cells, and chondrocytes [1, 2]
mesenchymal stem cells (MSC-ATs), which can be collected from AT, have been applied to the treatment of a wide range of diseases in light of the low invasiveness compared with surgery
The functions of proteins classified by the GO analysis were quantified using the liquid chromatograph (LC)-MS/MS measurement system for the amount of proteins and components contained in primary cultured cells of mMSC-ATs and cells passaged three times
Summary
Mesenchymal stromal stem cells (MSCs) are considered to have the ability to differentiate into mesenchymal cells, such as osteoblasts, adipocytes, muscle cells, and chondrocytes [1, 2]. One method of sterilizing tissues collected from a living body involves immersing and storing such tissue for 16 h using HBSS [13], which contains antibiotics After such storage, MSC-ATs can be isolated from adipose tissue. MSC-ATs of primary cultured cells are reportedly contaminated with various types of cells, such as blood cells, through the process of cell isolation This is because the stromal vascular fraction (SVF) [15] obtained when collecting MSC-ATs using centrifugation contains many kinds of cells (e.g., adipocytes, fibroblasts, smooth muscle cells, endothelial cells, blood cells, endothelial progenitor cells, preadipocytes, vascular progenitors, hematopoietic progenitors, and hematopoietic stem cells) [16, 17]. Determining the protein component of MSCATs that exerts a therapeutic effect is expected to be useful for cell therapy in the future
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