A Liquid Chromatography with Tandem Mass Spectrometry-Based Proteomic Analysis of Cells Cultured in DMEM 10% FBS and Chemically Defined Medium Using Human Adipose-Derived Mesenchymal Stem Cells.
Yoshiki NakashimaSaifun NaharChika Miyagi-ShiohiraTakao KinjoNaoya KobayashiIssei SaitohMasami WatanabeJiro FujitaHirofumi NoguchiPublished in: International journal of molecular sciences (2018)
Human adipose-derived mesenchymal stem cells (hADSCs) are representative cell sources for cell therapy. Classically, Dulbecco's Modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) has been used as culture medium for hADSCs. A chemically defined medium (CDM) containing no heterologous animal components has recently been used to produce therapeutic hADSCs. However, how the culture environment using a medium without FBS affects the protein expression of hADSC is unclear. We subjected hADSCs cultured in CDM and DMEM (10% FBS) to a protein expression analysis by tandem mass spectrometry liquid chromatography and noted 98.2% agreement in the proteins expressed by the CDM and DMEM groups. We classified 761 proteins expressed in both groups by their function in a gene ontology analysis. Thirty-one groups of proteins were classified as growth-related proteins in the CDM and DMEM groups, 16 were classified as antioxidant activity-related, 147 were classified as immune system process-related, 557 were involved in biological regulation, 493 were classified as metabolic process-related, and 407 were classified as related to stimulus responses. These results show that the trend in the expression of major proteins related to the therapeutic effect of hADSCs correlated strongly in both groups.
Keyphrases
- tandem mass spectrometry
- liquid chromatography
- cell therapy
- mass spectrometry
- simultaneous determination
- high performance liquid chromatography
- high resolution mass spectrometry
- poor prognosis
- type diabetes
- high resolution
- adipose tissue
- stem cells
- gene expression
- cell death
- single cell
- insulin resistance
- metabolic syndrome
- small molecule
- dna methylation
- mesenchymal stem cells
- cell proliferation
- pi k akt
- cell cycle arrest
- signaling pathway