Effects of culture conditions on osteogenic differentiation in human mesenchymal stem cells.
Human bone marrow-derived mesenchymal stem cells (hBMMSCs) must differentiate into osteogenic cells to allow for successful bone regeneration. In this study, we investigated the effects of different combinations of three soluble osteogenic differentiation-inducing factors [L-ascorbic acid (AC), beta-glycerophosphate (betaG), and bone morphogenic protein-2 (BMP-2)] and the presence of a hydroxyapatite (HA) substrate on hBMMSC osteogenic differentiation in vitro. hBMMSCs were cultured in medium containing various combinations of the soluble factors on culture plates with or without HA coating. After 7 days of culture, alkaline phosphatase (ALP) activity, calcium deposition, and osteoprotegerin (OPG) and osteopontin (OPN) expression were measured. The effects of individual and combined factors were evaluated using a factorial analysis method. BMP-2 predominantly affected expression of early markers of osteogenic differentiation (ALP and OPG). HA had the highest positive effect on OPN expression and calcium deposition. The interaction between AC, betaG, and HA had the second highest positive effect on ALP activity.
Song
Su Jin
SJ
Department of Chemical Engineering, Hanyang University, Seoul 133-791, Korea.
Jeon
Oju
O
Yang
Hee Seok
HS
Han
Dong Keun
DK
Kim
Byung-Soo
BS
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D001205:Ascorbic Acid / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible; D004305:Dose-Response Relationship, Drug; D017886:Durapatite / Q000494:pharmacology*; D005994:Glycerophosphates / Q000494:pharmacology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects*; D053495:Osteopontin / Q000032:analysis / Q000378:metabolism; D053244:Osteoprotegerin / Q000032:analysis / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D020099:Coated Materials, Biocompatible; D005994:Glycerophosphates; D053244:Osteoprotegerin; D016212:Transforming Growth Factor beta; D053495:Osteopontin; D017886:Durapatite; D000469:Alkaline Phosphatase; D001205:Ascorbic Acid; D002118:Calcium; C031463:beta-glycerophosphoric acid