Stable CpG Hypomethylation of Adipogenic Promoters in Freshly Isolated, Cultured, and Differentiated Mesenchymal Stem Cells from Adipose Tissue

Noer, Agate; Sørensen, Anita L.; Boquest, Andrew C.; Collas, Philippe

doi:10.1091/mbc.e06-04-0322

Cited by 130 publications

(188 citation statements)

References 73 publications

(84 reference statements)

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“…However, in our study, the methylation status of promoter CpG islands was found to be preserved in critical genes, such as SOX9 and RUNX2, during chondrogenesis. This observation may indicate a retained capability of the mesenchymal progenitor cells to change their differentiation status even after chondrogenesis, since similar observations about adipogenesis of MSCs have been reported (36)(37)(38)(39).…”

Section: Discussionsupporting

confidence: 82%

Methylation status of CpG islands in the promoter regions of signature genes during chondrogenesis of human synovium–derived mesenchymal stem cells

Ezura

Sekiya

Koga

et al. 2009

Arthritis & Rheumatism

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Objective. Human synovium-derived mesenchymal stem cells (MSCs) can efficiently differentiate into mature chondrocytes. It has been suggested that DNA methylation is one mechanism that regulates human chondrogenesis; however, the methylation status of genes related to chondrogenic differentiation is not known. The purpose of this study was to investigate the CpG methylation status in human synovium-derived MSCs during experimental chondrogenesis, with a view toward potential therapeutic use in osteoarthritis.Methods. Human synovium-derived MSCs were subjected to chondrogenic pellet culture for 3 weeks. The methylation status of 12 regions in the promoters of 10 candidate genes (SOX9, RUNX2, CHM1, FGFR3, CHAD, MATN4, SOX4, GREM1, GPR39, and SDF1) was analyzed by bisulfite sequencing before and after differentiation. The expression levels of these genes were analyzed by real-time reverse transcription-polymerase chain reaction. Methylation status was also examined in human articular cartilage.Results. Bisulfite sequencing analysis indicated that 10 of the 11 CpG-rich regions analyzed were hypomethylated in human progenitor cells before and after 3 weeks of pellet culture, regardless of the expression levels of the genes. The methylation status was consistently low in SOX9, RUNX2, CHM1, CHAD, and FGFR3 following an increase in expression upon differentiation and was low in GREM1 and GPR39 following a decrease in expression upon chondrogenesis. One exceptional instance of a differentially methylated CpGrich region was in a 1-kb upstream sequence of SDF1, the expression of which decreased upon differentiation. Paradoxically, the hypermethylation status of this region was reduced after 3 weeks of pellet culture.Conclusion. The DNA methylation levels of CpGrich promoters of genes related to chondrocyte phenotypes are largely kept low during chondrogenesis in human synovium-derived MSCs.

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Section: Discussionsupporting

confidence: 82%

Methylation status of CpG islands in the promoter regions of signature genes during chondrogenesis of human synovium–derived mesenchymal stem cells

Ezura

Sekiya

Koga

et al. 2009

Arthritis & Rheumatism

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“…44 Similarly, under 5-azacytidine treatment, commitment of 10T1/2 cells to the myogenic lineage correlates with DNA demethylation of the promoter region of the myogenic transcription factor myod gene. 45 In accordance with all these data, the promoters of adipogenic genes such as pparg2 or leptin are unmethylated in isolated adipose tissue stromal cells enriched in preadipocytes, whereas myogenic and endothelial promoters are methylated, 46 indicating that these cells have already committed to an adipogenic fate and additionally supporting a role for DNA methylation on lineage selection and cell differentiation. Moreover, increased levels of pparg2 promoter methylation have been described in visceral adipose tissue of several mouse models of diabetes 47 indicating that the gene needs to remain unmethylated in order to maintain the mature characteristics of the cells.…”

supporting

confidence: 69%

A chromatin perspective of adipogenesis

2010

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“…DNA methylation favors genomic integrity and ensures proper regulation of gene expression. Corruption of DNA methylation in long-term culture of primary cells, including ASCs (Noer et al, 2006;Boquest et al, 2007), are predominantly caused by stochastic CpG methylation events reflecting errors in the maintenance methylation machinery (Graff et al, 2000;Bird, 2002). DNA methylation is associated with longterm gene silencing (Antequera, 2003), thus methylation of tumor suppressor genes may lead to cellular transformation (Laird, 2005).…”

Section: Introductionmentioning

confidence: 99%

Genetic and epigenetic instability of human bone marrow mesenchymal stem cells expanded in autologous serum or fetal bovine serum

Dahl

Duggal²,

Coulston³

et al. 2008

Int. J. Dev. Biol.

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163

119

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Culture of mesenchymal stem cells (MSCs) under conditions promoting proliferation and differentiation, while supporting genomic and epigenetic stability, is essential for therapeutic use. We report here the extent of genome-wide DNA gains and losses and of DNA methylation instability on 170 cancer-related promoters in bone marrow (BM) MSCs during culture to late passage in medium containing fetal bovine serum (FBS) or autologous serum (AS). Comparative genomic hybridization indicates that expansion of BMMSCs elicits primarily telomeric deletions in a subpopulation of cells, the extent of which varies between donors. However, late passage cultures in AS consistently display normal DNA copy numbers. Combined bisulfite restriction analysis and bisulfite sequencing show that although DNA methylation states are overall stable in culture, AS exhibits stronger propensity than FBS to maintain unmethylated states. Comparison of DNA methylation in BMMSCs with freshly isolated and cultured adipose stem cells (ASCs) also reveals that most genes unmethylated in both BMMSCs and ASCs in early passage are also unmethylated in uncultured ASCs. We conclude that (i) BMMSCs expanded in AS or FBS may display localized genetic alterations, (ii) AS tends to generate more consistent genomic backgrounds and DNA methylation patterns, and (iii) the unmethylated state of uncultured MSCs is more likely to be maintained in culture than the methylated state.

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Stable CpG Hypomethylation of Adipogenic Promoters in Freshly Isolated, Cultured, and Differentiated Mesenchymal Stem Cells from Adipose Tissue

Cited by 130 publications

References 73 publications

Methylation status of CpG islands in the promoter regions of signature genes during chondrogenesis of human synovium–derived mesenchymal stem cells

Methylation status of CpG islands in the promoter regions of signature genes during chondrogenesis of human synovium–derived mesenchymal stem cells

A chromatin perspective of adipogenesis

Genetic and epigenetic instability of human bone marrow mesenchymal stem cells expanded in autologous serum or fetal bovine serum

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