Paweena Wongwitwichot1, Jasadee Kaewsrichan2. 1. Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat-yai, Songkhla, Thailand. 2. Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences and Nanotec-PSU Center of Excellence on Drug Delivery System, Prince of Songkla University, Hat-yai, Songkhla, Thailand. Electronic address: jasadee.k@psu.ac.th.
Abstract
PURPOSE: Mesenchymal stem cells (MSCs) are multipotent adult stem cells and present in practically all tissues but originally identified within the bone marrow (BM). The differentiation potential of these cells is generally impaired when culturing in vitro for cell expansion. The aim of this study is to speedily increase the numbers of bone marrow derived mesenchymal stem cells (BM-MSCs) with substantially maintaining their differentiation potential in vitro and improving bone formation in vivo. MATERIALS AND METHODS: BM-MSCs isolated from rats were sequentially cultured in α-MEM containing basic fibroblast growth factor (FGF2) and/or insulin to stimulate proliferation and osteogenic commitment, and in the medium with the addition of bone morphogenetic protein 2 (BMP2) and/or bone morphogenetic protein 7 (BMP7) to arouse differentiation. The expression of genes markedly associating the commitment and differentiation were investigated in vitro using real-time PCR technique and mineralization assay, while the capacity of inducing bone formation by the established conditions was determined in vivo using a rat model. RESULTS: The BM-MSCs greatly proliferated with active transcription of runx2 and osterix genes when induced by FGF2 and insulin. The in vitro mineralization was enhanced by BMP2, but the extent was diminished when BMP2 was replaced or supplemented by BMP7. Formation of new small blood vessels was notably detected when the cells were respectively challenged by FGF2 plus insulin and BMP2. CONCLUSION: These data are valuable in choosing growth factors for proper bone repair. However, optimization of the established system would be essential when the cells of human source are applied.
PURPOSE: Mesenchymal stem cells (MSCs) are multipotent adult stem cells and present in practically all tissues but originally identified within the bone marrow (BM). The differentiation potential of these cells is generally impaired when culturing in vitro for cell expansion. The aim of this study is to speedily increase the numbers of bone marrow derived mesenchymal stem cells (BM-MSCs) with substantially maintaining their differentiation potential in vitro and improving bone formation in vivo. MATERIALS AND METHODS: BM-MSCs isolated from rats were sequentially cultured in α-MEM containing basic fibroblast growth factor (FGF2) and/or insulin to stimulate proliferation and osteogenic commitment, and in the medium with the addition of bone morphogenetic protein 2 (BMP2) and/or bone morphogenetic protein 7 (BMP7) to arouse differentiation. The expression of genes markedly associating the commitment and differentiation were investigated in vitro using real-time PCR technique and mineralization assay, while the capacity of inducing bone formation by the established conditions was determined in vivo using a rat model. RESULTS: The BM-MSCs greatly proliferated with active transcription of runx2 and osterix genes when induced by FGF2 and insulin. The in vitro mineralization was enhanced by BMP2, but the extent was diminished when BMP2 was replaced or supplemented by BMP7. Formation of new small blood vessels was notably detected when the cells were respectively challenged by FGF2 plus insulin and BMP2. CONCLUSION: These data are valuable in choosing growth factors for proper bone repair. However, optimization of the established system would be essential when the cells of human source are applied.