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Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes.

Lee HM, Joo BS, Lee CH, Kim HY, Ock JH, Lee YS - J Menopausal Med (2015)

Bottom Line: In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2).This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Natural Science College, Pusan National University, Busan, Korea.

ABSTRACT

Objectives: Glucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes.

Methods: ADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined.

Results: Isolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).

Conclusion: This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

No MeSH data available.


Characterization of adipose-derived stem cells (ADSCs). (A) Flow cytometry analysis of ADSCs. Cells were stained with fluorescein fluorescein isothiocyanate (FITC)-labeled CD73, CD90, CD105, CD31, CD34 and CD45 antibodies. Fluorochrome-conjugated nonspecific mouse IgG1 was used as isotype controls (dotted red lines). ADSCs were stained positively for surface markers characteristic of mesenchymal stem cell, CD73, CD90 and CD105. The cells were negative for markers of the endothelial cells CD31, hematopoietic lineage CD45 and of hematopoietic stem cells CD34. (B, C) Osteogenic differentiation of ADSCs. Seven days of osteogenic induction, mRNA expressions of osteogenic markers were observed by reverse transcriptase-polymerase chain reaction (RT-PCR). Fourteen days of osteogenic induction, the deposition of calcium precipitates was stained by alizarin red S staining. (D, E) Adipogenic differentiation of ADSCs. Seven days of adipogenic induction, mRNA expressions of adipocyte markers were detected by RT-PCR. Fourteen days of adipogenic induction, the lipid droplets were stained by Oil red O staining. Original magnification, × 100. Original magnification, × 100. FITC: fluorescein isothiocyanate, OIM: osteogenic induction media, ALP: alkaline phosphatase, RUNX2: runt-related transcription factor 2, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, AIM: adipogenic induction media, AP2: adipocyte protein 2, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor-gamma.
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Figure 1: Characterization of adipose-derived stem cells (ADSCs). (A) Flow cytometry analysis of ADSCs. Cells were stained with fluorescein fluorescein isothiocyanate (FITC)-labeled CD73, CD90, CD105, CD31, CD34 and CD45 antibodies. Fluorochrome-conjugated nonspecific mouse IgG1 was used as isotype controls (dotted red lines). ADSCs were stained positively for surface markers characteristic of mesenchymal stem cell, CD73, CD90 and CD105. The cells were negative for markers of the endothelial cells CD31, hematopoietic lineage CD45 and of hematopoietic stem cells CD34. (B, C) Osteogenic differentiation of ADSCs. Seven days of osteogenic induction, mRNA expressions of osteogenic markers were observed by reverse transcriptase-polymerase chain reaction (RT-PCR). Fourteen days of osteogenic induction, the deposition of calcium precipitates was stained by alizarin red S staining. (D, E) Adipogenic differentiation of ADSCs. Seven days of adipogenic induction, mRNA expressions of adipocyte markers were detected by RT-PCR. Fourteen days of adipogenic induction, the lipid droplets were stained by Oil red O staining. Original magnification, × 100. Original magnification, × 100. FITC: fluorescein isothiocyanate, OIM: osteogenic induction media, ALP: alkaline phosphatase, RUNX2: runt-related transcription factor 2, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, AIM: adipogenic induction media, AP2: adipocyte protein 2, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor-gamma.

Mentions: In order to assess whether ADSCs have MSC-specific characteristics, flow cytometry was performed at passage 4. The cells were positively stained for MSC-specific surface markers (CD73, CD90 and CD105), but they were negative for markers of the endothelial cells (CD31) and hematopoietic cells (CD34 and CD45) (Fig. 1A).


Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes.

Lee HM, Joo BS, Lee CH, Kim HY, Ock JH, Lee YS - J Menopausal Med (2015)

Characterization of adipose-derived stem cells (ADSCs). (A) Flow cytometry analysis of ADSCs. Cells were stained with fluorescein fluorescein isothiocyanate (FITC)-labeled CD73, CD90, CD105, CD31, CD34 and CD45 antibodies. Fluorochrome-conjugated nonspecific mouse IgG1 was used as isotype controls (dotted red lines). ADSCs were stained positively for surface markers characteristic of mesenchymal stem cell, CD73, CD90 and CD105. The cells were negative for markers of the endothelial cells CD31, hematopoietic lineage CD45 and of hematopoietic stem cells CD34. (B, C) Osteogenic differentiation of ADSCs. Seven days of osteogenic induction, mRNA expressions of osteogenic markers were observed by reverse transcriptase-polymerase chain reaction (RT-PCR). Fourteen days of osteogenic induction, the deposition of calcium precipitates was stained by alizarin red S staining. (D, E) Adipogenic differentiation of ADSCs. Seven days of adipogenic induction, mRNA expressions of adipocyte markers were detected by RT-PCR. Fourteen days of adipogenic induction, the lipid droplets were stained by Oil red O staining. Original magnification, × 100. Original magnification, × 100. FITC: fluorescein isothiocyanate, OIM: osteogenic induction media, ALP: alkaline phosphatase, RUNX2: runt-related transcription factor 2, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, AIM: adipogenic induction media, AP2: adipocyte protein 2, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor-gamma.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 1: Characterization of adipose-derived stem cells (ADSCs). (A) Flow cytometry analysis of ADSCs. Cells were stained with fluorescein fluorescein isothiocyanate (FITC)-labeled CD73, CD90, CD105, CD31, CD34 and CD45 antibodies. Fluorochrome-conjugated nonspecific mouse IgG1 was used as isotype controls (dotted red lines). ADSCs were stained positively for surface markers characteristic of mesenchymal stem cell, CD73, CD90 and CD105. The cells were negative for markers of the endothelial cells CD31, hematopoietic lineage CD45 and of hematopoietic stem cells CD34. (B, C) Osteogenic differentiation of ADSCs. Seven days of osteogenic induction, mRNA expressions of osteogenic markers were observed by reverse transcriptase-polymerase chain reaction (RT-PCR). Fourteen days of osteogenic induction, the deposition of calcium precipitates was stained by alizarin red S staining. (D, E) Adipogenic differentiation of ADSCs. Seven days of adipogenic induction, mRNA expressions of adipocyte markers were detected by RT-PCR. Fourteen days of adipogenic induction, the lipid droplets were stained by Oil red O staining. Original magnification, × 100. Original magnification, × 100. FITC: fluorescein isothiocyanate, OIM: osteogenic induction media, ALP: alkaline phosphatase, RUNX2: runt-related transcription factor 2, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, AIM: adipogenic induction media, AP2: adipocyte protein 2, LPL: lipoprotein lipase, PPAR-γ: peroxisome proliferator-activated receptor-gamma.
Mentions: In order to assess whether ADSCs have MSC-specific characteristics, flow cytometry was performed at passage 4. The cells were positively stained for MSC-specific surface markers (CD73, CD90 and CD105), but they were negative for markers of the endothelial cells (CD31) and hematopoietic cells (CD34 and CD45) (Fig. 1A).

Bottom Line: In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2).This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Natural Science College, Pusan National University, Busan, Korea.

ABSTRACT

Objectives: Glucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes.

Methods: ADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined.

Results: Isolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-γ), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05).

Conclusion: This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

No MeSH data available.