title: Optimized osteogenic differentiation protocol from R1 mouse embryonic stem cells in vitro.
abstract: Embryonic stem cells (ESCs) are a unique model that allows the study of molecular pathways underlying commitment and differentiation. One such lineage is osteoblasts, which are responsible for forming bone tissue in the body. There are many osteogenic differentiation protocols in the literature utilizing different soluble factors. The goal of the present study was to increase the efficacy of our osteogenic differentiation protocol from R1 cells. We have studied the effects of the addition of the following factors: dexamethasone, retinoic acid, and peroxisome-proliferator-activated receptor-gamma inhibitor, which have been reported to enhance osteogenesis. We found that among the 6 different protocols that were tested, the addition of retinoic acid with later addition of dexamethasone gives the most enrichment of osteogenic lineage cells. Thus, our findings provide valuable guidelines for culture condition to differentiate mouse R1 ESCs to osteoblastic cells in vitro.
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, M5S 1A8 Canada.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, M5S 1A8 Canada; Department of Chemistry & Earth Sciences, College of Arts and Science, University of Qatar, P.O. Box 2713, Doha, Qatar.
authors.forename: Layla
authors.initials: L
authors.lastname: Al-Mansoori
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, M5S 1A8 Canada. Electronic address: m.opas@utoronto.ca.
authors.forename: Michal
authors.initials: M
authors.lastname: Opas
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D019070:Cell Lineage; D003907:Dexamethasone / Q000008:administration & dosage; D066298:In Vitro Techniques / Q000379:methods*; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000187:drug effects*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics*; D047495:PPAR gamma / Q000037:antagonists & inhibitors; D014212:Tretinoin / Q000008:administration & dosage
chemical_list: D047495:PPAR gamma; D014212:Tretinoin; D003907:Dexamethasone
keywords: Embryonic stem cells; In vitro culture; Osteogenic differentiation