Endocrinol Metab.  2014 Dec;29(4):567-573. 10.3803/EnM.2014.29.4.567.

Expression of Glucagon-Like Peptide 1 Receptor during Osteogenic Differentiation of Adipose-Derived Stem Cells

• Jeon YK
• Bae MJ
• Kim JI
• Kim JH
• Choi SJ
• Kwon SK
• An JH
• Kim SS
• Kim BH
• Kim YK
• Kim IJ

• KMID: 2384256
• DOI: http://doi.org/10.3803/EnM.2014.29.4.567

Abstract

BACKGROUND
Glucagon-like peptide 1 (GLP-1), an incretin hormone well known for its glucose-lowering effect, was recently reported to exert an anabolic effect on bone. Although the exact mechanism is not known, it likely involves the GLP-1 receptor (GLP-1R), which is expressed in some osteoblastic cell lines. Adipose-derived stem cells (ADSCs) have mesenchymal stem cell-specific characteristics, including osteoblastic differentiation potential. We evaluated the expression of GLP-1R during osteogenic differentiation of ADSCs.
METHODS
ADSCs were isolated from subcutaneous adipose tissue obtained from three male donors during plastic surgery and were subjected to osteogenic induction. Mineralization was assessed by Alizarin Red staining on day 21. Expression of alkaline phosphatase (ALP), osteocalcin (OC), and GLP-1R was measured by real-time polymerase chain reaction in triplicate for each patient on days 0, 7, 14, and 21. Target mRNA expression levels were normalized to that of beta-actin.
RESULTS
ADSCs were fibroblast-like in morphology, adhered to plastic, and had multipotent differentiation potential, as assessed using specific antigen markers. The osteogenic markers ALP and OC were notably upregulated at 21 days. Osteogenic differentiation resulted in a time-dependent increase in the expression of GLP-1R (P=0.013).
CONCLUSION
We demonstrated upregulation of GLP-1R gene expression during osteogenic differentiation of ADSCs. This finding suggests that GLP-1 may induce osteogenic differentiation in bone tissue.

Keyword

Glucagon-like peptide 1; Glucagon-like peptide 1 receptor; Adipose-derived stem cell; Osteogenesis

MeSH Terms

Actins
Alkaline Phosphatase
Anabolic Agents
Bone and Bones
Cell Line
Gene Expression
Glucagon-Like Peptide 1*
Humans
Incretins
Male
Osteoblasts
Osteocalcin
Osteogenesis
Real-Time Polymerase Chain Reaction
RNA, Messenger
Stem Cells*
Subcutaneous Fat
Surgery, Plastic
Tissue Donors
Up-Regulation
Glucagon-Like Peptide-1 Receptor
Actins
Alkaline Phosphatase
Anabolic Agents
Glucagon-Like Peptide 1
Incretins
Osteocalcin
RNA, Messenger

Figure

• 

  Fig. 1 Adipose-derived stem cells (ADSCs). (A) ADSCs exhibited fibroblast-like morphology and adhered to plastic, as assessed by light microscopy (×40) at passage three. (B) Flow cytometry using mesenchyme-specific antigen markers showed that ADSCs express CD73, CD90, and CD105, but not CD11b, CD19, CD31, and HLA-DR.

• 

  Fig. 2 Osteogenic differentiation of adipose-derived stem cells. (A) Calcification, detected by Alizarin red staining and indicative of mineralization, was assessed by (Aa) gross appearance and (Ab) light microscopy (×40) at days 0, 7, 14, and 21. Real-time polymerase chain reaction showed increased expression of (B) alkaline phosphatase (ALP) and (C) osteocalcin mRNA at day 21. Data are presented as means±standard deviation.

• 

  Fig. 3 Expression of glucagon-like peptide 1 receptor (GLP-1R) during osteogenic differentiation of adipose-derived stem cells (ADSCs). (A) Western blotting for GLP-1R in differentiating ADSCs at days 0, 7, 14, and 21 after differentiation. Data are presented as means±standard deviation. (B) The expression level of GLP-1R mRNA increased significantly in a time-dependent manner during osteogenic differentiation (P=0.013). GAPDH, glyceraldehyde 3-phosphate dehydrogenase. aP<0.05.

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