title: Development of a rapid culture method to induce adipocyte differentiation of human bone marrow-derived mesenchymal stem cells.
abstract: Human mesenchymal stem cells (hMSCs) derived from bone marrow are multipotent stem cells that can regenerate mesenchymal tissues such as adipose, bone or muscle. It is thought that hMSCs can be utilized as a cell resource for tissue engineering and as human models to study cell differentiation mechanisms, such as adipogenesis, osteoblastogenesis and so on. Since it takes 2-3weeks for hMSCs to differentiate into adipocytes using conventional culture methods, the development of methods to induce faster differentiation into adipocytes is required. In this study we optimized the culture conditions for adipocyte induction to achieve a shorter cultivation time for the induction of adipocyte differentiation in bone marrow-derived hMSCs. Briefly, we used a cocktail of dexamethasone, insulin, methylisobutylxanthine (DIM) plus a peroxisome proliferator-activated receptor gamma agonist, rosiglitazone (DIMRo) as a new adipogenic differentiation medium. We successfully shortened the period of cultivation to 7-8days from 2-3weeks. We also found that rosiglitazone alone was unable to induce adipocyte differentiation from hMSCs in vitro. However, rosiglitazone appears to enhance hMSC adipogenesis in the presence of other hormones and/or compounds, such as DIM. Furthermore, the inhibitory activity of TGF-beta1 on adipogenesis could be investigated using DIMRo-treated hMSCs. We conclude that our rapid new culture method is very useful in measuring the effect of molecules that affect adipogenesis in hMSCs.
authors.affiliation: Translational Research Center, Saitama International Medical, Saitama Medical University, 1397-1 Yamane, Hidaka, Saitama 350-1298, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Ninomiya
authors.forename: Yzumi
authors.initials: Y
authors.lastname: Sugahara-Yamashita
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Nakachi
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Tokuzawa
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okazaki
authors.forename: Masahiko
authors.initials: M
authors.lastname: Nishiyama
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D017667:Adipocytes / Q000166:cytology*; D050156:Adipogenesis; D018929:Cell Culture Techniques; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D050356:Lipid Metabolism / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: D003470:Culture Media; D007328:Insulin; D045162:Thiazolidinediones; D016212:Transforming Growth Factor beta; D000077154:Rosiglitazone; D003907:Dexamethasone; D015056:1-Methyl-3-isobutylxanthine