Gene Expression Pattern of Human Chorion-Derived Mesenchymal Stem Cells during Adipogenic Differentiation
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: The aim of this study was to identify the adipocyte-specific gene expression patterns in chorion-derived mesenchymal stem cells during adipogenic differentiation. Materials and Methods: Chorionic cells were isolated from the third trimester chorions from human placenta at birth and identified morphologically and by fluorescence-activated cell sorting analysis. After inducing adipogenic differentiation for 28 days, cells at days 3, 10, 21 and 28 were analyzed by Oil red O staining and RNA extraction in order to assess the expression levels of adipocyte marker genes, including CCAAT-enhancer binding protein α (C/EBPα), peroxisome proliferator-activated receptor γ (PPARγ), fatty acid binding protein 4 (FABP4) and Glycerol-3-phosphate dehydrogenase (GPD2). Cells not induced for differentiation were compared with the induced cells as a control group. Results: Chorion-derived cells showed the same pattern as fibroblasts, and expressed CD73, CD105, and CD166 antigens, but not CD45, CD34, and HLA-DR antigens. On day 3 after differentiation,cells began to stain positively upon Oil red O staining, and continuously increased in lipid granules for 4 weeks. The expression level of C/EBPα increased 4.6 fold on day 3 after induction, and continued to increase for 4 weeks. PPARγ was expressed at a maximum of 2.9 fold on day 21. FABP4 and GPD2 were significantly expressed at 4.7- and 3.0-fold, respectively, on day 21, compared to controls, and further increased thereafter. Conclusion: Human chorion-derived mesenchymal stem cells exhibited the sequential expression pattern of adipocyte marker genes during differentiation, corresponding to adipogenesis.
Abstract
Purpose: The aim of this study was to identify the adipocyte-specific gene expression patterns in chorion-derived mesenchymal stem cells during adipogenic differentiation. Materials and Methods: Chorionic cells were isolated from the third trimester chorions from human placenta at birth and identified morphologically and by fluorescence-activated cell sorting analysis. After inducing adipogenic differentiation for 28 days, cells at days 3, 10, 21 and 28 were analyzed by Oil red O staining and RNA extraction in order to assess the expression levels of adipocyte marker genes, including CCAAT-enhancer binding protein α (C/EBPα), peroxisome proliferator-activated receptor γ (PPARγ), fatty acid binding protein 4 (FABP4) and Glycerol-3-phosphate dehydrogenase (GPD2). Cells not induced for differentiation were compared with the induced cells as a control group. Results: Chorion-derived cells showed the same pattern as fibroblasts, and expressed CD73, CD105, and CD166 antigens, but not CD45, CD34, and HLA-DR antigens. On day 3 after differentiation,cells began to stain positively upon Oil red O staining, and continuously increased in lipid granules for 4 weeks. The expression level of C/EBPα increased 4.6 fold on day 3 after induction, and continued to increase for 4 weeks. PPARγ was expressed at a maximum of 2.9 fold on day 21. FABP4 and GPD2 were significantly expressed at 4.7- and 3.0-fold, respectively, on day 21, compared to controls, and further increased thereafter. Conclusion: Human chorion-derived mesenchymal stem cells exhibited the sequential expression pattern of adipocyte marker genes during differentiation, corresponding to adipogenesis.
참고문헌 (30)
- Pittenger MF, "Multilineage potential of adult human mesenchymal stem cells", Science, 284, pp.143~147 (1999)
- Ferrari G, "Muscle regeneration by bone marrow-derived myogenic progenitors", Science, 279, pp.1528~1530 (1998)
- Sanchez-Ramos J, "Adult bone marrow stromal cells differentiate into neural cells in vitro", Exp Neurol, 164, pp.247~256 (2000)
- Beyer Nardi N, "Mesenchymal stem cells: isolation, in vitro expansion and characterization", Handb Exp Pharmacol, pp.249~282 (2006)
- Dvash T, "Temporal gene expression during differentiation of human embryonic stem cells and embryoid bodies", Hum Reprod, 19, pp.2875~2883 (2004)
- Yamamoto Y, "A comparative analysis of the transcriptome and signal pathways in hepatic differentiation of human adipose mesenchymal stem cells", FEBS J, 275, pp.1260~1273 (2008)
- Wang Q, "Temporal expression of estrogen receptor alpha in rat bone marrow mesenchymal stem cells", Biochem Biophys Res Commun, 347, pp.117~123 (2006)
- Hu Y, "Isolation and identification of mesenchymal stem cells from human fetal pancreas", J Lab Clin Med, 141, pp.342~349 (2003)
- in ‘t Anker PS, "Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential", Haematologica, 88, pp.845~852 (2003)
- Campagnoli C, "Identification of mesenchymal stem", progenitor cells in human first-trimester fetal blood, liver, and bone marrow, 2001, pp.98, 2396~2402
- Battula VL, "Human placenta and bone marrow derived MSC cultured in serum-free, b-FGF-containing medium express cell surface frizzled-9 and SSEA-4 and give rise to multilineage differentiation", Differentiation, 75, pp.279~291 (2007)
- In ‘t Anker PS, "Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta", Stem Cells, 22, pp.1338~1345 (2004)
- Miki T, "Stem cell characteristics of amniotic epithelial cells", Stem Cells, 23, pp.1549~1559 (2005)
- Mitchell KE, "Matrix cells from Wharton’s jelly form neurons and glia", Stem Cells, 21, pp.50~60 (2003)
- Portmann-Lanz CB, "Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration", Am J Obstet Gynecol, 194, pp.664~673 (2006)
- Otto TC, "Adipose development: from stem cell to adipocyte", Crit Rev Biochem Mol Biol, 40, pp.229~242 (2005)
- Hutley LJ, "Effects of rosiglitazone and linoleic acid on human preadipocyte differentiation", Eur J Clin Invest, 33, pp.574~581 (2003)
- Soncini M, "Isolation and characterization of mesenchymal cells from human fetal membranes", J Tissue Eng Regen Med, 1, pp.296~305 (2007)
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method", Methods, 25, pp.402~408 (2001)
- Umek RM, "CCAAT-enhancer binding protein: a component of a differentiation switch", Science, 251, pp.288~292 (1991)
- Tontonoz P, "mPPAR gamma 2: tissue-specific regulator of an adipocyte enhancer", Genes Dev, 8, pp.1224~1234 (1994)
- Rosen ED, "C", EBPalpha induces adipogenesis through PPARgamma: a unified pathway, 2002, pp.16, 22~26
- Shao D, "Peroxisome proliferator activated receptor gamma, CCAAT", enhancer-binding protein alpha, and cell cycle status regulate the commitment to adipocyte differentiation, 1997, pp.272, 21473~21478
- Wu Z, "Cross-regulation of C", EBP alpha and PPAR gamma controls the transcriptional pathway of adipogenesis and insulin sensitivity, 1999, pp.3, 151~158
- Auwerx J, "Transcription, adipocyte differentiation, and obesity", J Mol Med, 74, pp.347~352 (1996)
- Shin DW, "(-)-Catechin promotes adipocyte differentiation in human bone marrow mesenchymal stem cells through PPAR gamma transactivation", Biochem Pharmacol, 77, pp.125~133 (2009)
- Chang YJ, "Disparate mesenchyme-lineage tendencies in mesenchymal stem cells from human bone marrow and umbilical cord blood", Stem Cells, 24, pp.679~685 (2006)
- Karahuseyinoglu S, "Functional structure of adipocytes differentiated from human umbilical cord stroma-derived stem cells", Stem Cells, 26, pp.682~691 (2008)
- Sekiya I, "Adipogenic differentiation of human adult stem cells from bone marrow stroma (MSCs)", J Bone Miner Res, 19, pp.256~264 (2004)
- 김희경, "사람유래 지방줄기세포와 양수줄기세포의 분리 및 지방세포로의 분화에 따른 특성 비교", Obstetrics & Gynecology Science, 52(4), pp.447~455 (2009), 대한산부인과학회
