Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach.
Today, the tool that is most commonly used to evaluate the osteogenic differentiation of bone marrow stromal cells (BMSCs) in vitro is the demonstration of the expression of multiple relevant markers, such as ALP, RUNX2 and OCN. However, as yet, there is no single surface marker or panel of markers which clearly defines human BMSCs (hBMSCs) differentiating towards the osteogenic lineage. The aim of this study was therefore to examine this issue. Stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomics was utilized to investigate differently expressed surface markers in osteogenically differentiated and undifferentiated hBMSCs. Labeled membrane proteins were analyzed by mass spectrometry (MS) and 52 proteins with an expression ratio above 2, between osteogenically differentiated and undifferentiated cells, were identified. Subsequent validation, by flow cytometry and ELISA, of the SILAC expression ratios for a number of these proteins and investigations of the lineage specificity of three candidate markers were performed. The surface markers, CD10 and CD92, demonstrated significantly increased expression in hBMSCs differentiated towards the osteogenic and adipogenic lineages. In addition, there was a slight increase in CD10 expression during chondrogenic differentiation. Furthermore, the expression of the intracellular protein, crystalline-αB (CRYaB), was only significantly increased in osteogenically differentiated hBMSCs and not affected during differentiation towards the chondrogenic or adipogenic lineages. It has been concluded from the present results that CD10 and CD92 are potential markers of osteogenic and adipogenic differentiation and that CRYaB is a potential novel osteogenic marker specifically expressed during the osteogenic differentiation of hBMSCs in vitro.
Department of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden; BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden. Electronic address: cecilia.graneli@gu.se.
Cecilia
C
Granéli
Department of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden; BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden. Electronic address: anna.thorfve@biomaterials.gu.se.
Anna
A
Thorfve
Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy at The University of Gothenburg, Sweden. Electronic address: ulla.ruetschi@clinchem.gu.se.
Ulla
U
Ruetschi
BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden; Department of Orthopaedics, Sahlgrenska Academy at the University of Gothenburg, Sweden.
Helena
H
Brisby
Department of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden; BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden. Electronic address: peter.thomsen@biomaterials.gu.se.
Peter
P
Thomsen
BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden; Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy at The University of Gothenburg, Sweden. Electronic address: anders.lindahl@clinchem.gu.se.
Anders
A
Lindahl
Department of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden; BIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, Sweden. Electronic address: camilla.karlsson@biomaterials.gu.se.
Camilla
C
Karlsson
D050156:Adipogenesis; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis*; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007553:Isotope Labeling; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D015260:Neprilysin / Q000378:metabolism; D027701:Organic Cation Transport Proteins / Q000378:metabolism; D010012:Osteogenesis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D038203:alpha-Crystallin B Chain / Q000378:metabolism
D015703:Antigens, CD; D015415:Biomarkers; C059215:CRYAB protein, human; D027701:Organic Cation Transport Proteins; D020543:Proteome; C510732:SLC44A1 protein, human; D038203:alpha-Crystallin B Chain; D015260:Neprilysin