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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-01-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506113001359 |
| _version_ | 1818543190678962176 |
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| author | Cecilia Granéli Anna Thorfve Ulla Ruetschi Helena Brisby Peter Thomsen Anders Lindahl Camilla Karlsson |
| author_facet | Cecilia Granéli Anna Thorfve Ulla Ruetschi Helena Brisby Peter Thomsen Anders Lindahl Camilla Karlsson |
| author_sort | Cecilia Granéli |
| collection | DOAJ |
| description | 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. |
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| format | Article |
| id | doaj.art-89f802ccf5b7420582cc519f398637f7 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 1876-7753 |
| language | English |
| last_indexed | 2024-12-11T22:32:05Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-89f802ccf5b7420582cc519f398637f72022-12-22T00:48:05ZengElsevierStem Cell Research1873-50611876-77532014-01-0112115316510.1016/j.scr.2013.09.009Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approachCecilia Granéli0Anna Thorfve1Ulla Ruetschi2Helena Brisby3Peter Thomsen4Anders Lindahl5Camilla Karlsson6Department of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, SwedenDepartment of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy at The University of Gothenburg, SwedenBIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, SwedenDepartment of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, SwedenBIOMATCELL, VINN Excellence Center of Biomaterials and Cell Therapy, Gothenburg, SwedenDepartment of Biomaterials, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, SwedenToday, 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.http://www.sciencedirect.com/science/article/pii/S1873506113001359 |
| spellingShingle | Cecilia Granéli Anna Thorfve Ulla Ruetschi Helena Brisby Peter Thomsen Anders Lindahl Camilla Karlsson Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach Stem Cell Research |
| title | Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| title_full | Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| title_fullStr | Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| title_full_unstemmed | Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| title_short | Novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| title_sort | novel markers of osteogenic and adipogenic differentiation of human bone marrow stromal cells identified using a quantitative proteomics approach |
| url | http://www.sciencedirect.com/science/article/pii/S1873506113001359 |
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