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PMID:21401405

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Citation

Kim, HJ and Im, GI (2011) Electroporation-mediated transfer of SOX trio genes (SOX-5, SOX-6, and SOX-9) to enhance the chondrogenesis of mesenchymal stem cells. Stem Cells Dev. 20:2103-14

Abstract

The purpose of this study was to test the hypothesis that the SOX trio genes (SOX-5, SOX-6, and SOX-9) have a lower level of expression during the chondrogenic differentiation of mesenchymal stem cells (MSCs) compared with chondrocytes and that the electroporation-mediated gene transfer of SOX trio promotes chondrogenesis from human MSCs. An in vitro pellet culture was carried out using MSCs or chondrocytes at passage 3 and analyzed after 7 and 21 days. Then, MSCs were transfected with SOX trio genes and analyzed for the expression of chondrogenic markers after 21 days of in vitro culture. Without transforming growth factor-β1, the untransfected MSCs had a lower level of SOX trio gene and protein expression than chondrocytes. However, the level of SOX-9 gene expression increased in MSCs when treated with transforming growth factor-β1. GAG level significantly increased 7-fold in MSCs co-transfected with SOX trio, which was corroborated by Safranin-O staining. SOX trio co-transfection significantly increased COL2A1 gene and protein and decreased COL10A1 protein in MSCs. It is concluded that the SOX trio have a significantly lower expression in human MSCs than in chondrocytes and that the electroporation-mediated co-transfection of SOX trio enhances chondrogenesis and suppresses hypertrophy of human MSCs.

Links

PubMed Online version:10.1089/scd.2010.0516

Keywords

Aged; Apoptosis/genetics; Biological Markers/metabolism; Cell Count; Cell Proliferation; Chondrocytes/cytology; Chondrocytes/metabolism; Chondrogenesis/genetics; Collagen/genetics; Collagen/metabolism; DNA/metabolism; Electroporation/methods; Gene Expression Regulation; Glycosaminoglycans/metabolism; Green Fluorescent Proteins/metabolism; Humans; Mesenchymal Stem Cells/cytology; Mesenchymal Stem Cells/metabolism; Plasmids/genetics; Real-Time Polymerase Chain Reaction; SOX Transcription Factors/genetics; SOX Transcription Factors/metabolism; SOX9 Transcription Factor/genetics; SOX9 Transcription Factor/metabolism; SOXD Transcription Factors/genetics; SOXD Transcription Factors/metabolism; Transfection/methods

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:SOX5

involved_in

GO:0032332: positive regulation of chondrocyte differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX5

involved_in

GO:2000741: positive regulation of mesenchymal stem cell differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX5

involved_in

GO:0061036: positive regulation of cartilage development

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX5

involved_in

GO:0071560: cellular response to transforming growth factor beta stimulus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX6

involved_in

GO:0032332: positive regulation of chondrocyte differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX6

involved_in

GO:2000741: positive regulation of mesenchymal stem cell differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX6

involved_in

GO:0061036: positive regulation of cartilage development

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX6

involved_in

GO:0071560: cellular response to transforming growth factor beta stimulus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX9

involved_in

GO:0032332: positive regulation of chondrocyte differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX9

involved_in

GO:0061036: positive regulation of cartilage development

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX9

involved_in

GO:0071560: cellular response to transforming growth factor beta stimulus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:SOX9

involved_in

GO:2000741: positive regulation of mesenchymal stem cell differentiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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