title: Osteogenic differentiation strategies for adipose-derived mesenchymal stem cells.
abstract: Adipose stem cell preparations, either obtained as a freshly isolated so-called stromal vascular fraction (SVF) or as cells cultured to homogeneity and then referred to as adipose stem cells (ASCs), have found widespread use in a broad variety of studies on tissue engineering and regenerative medicine applications, including bone repair.For newcomers within the field, but also for established research laboratories having up to 10 years of expertise in this research area, it may be convenient to strive for, and use consensus protocols (1) for studying the osteogenic differentiation potential of ASC preparations in vitro, and (2) for osteogenic induction regimes for in vivo implementation. To assist in achieving this goal, this chapter describes various step-by-step osteogenic differentiation protocols for adipose-derived stem cell populations (SVF as well as ASCs) currently applied within our laboratory, with particular emphasis on protocols aimed at intra-operative use. The protocols describe the use of inducing compounds, including the bone morphogenetic proteins (BMPs), 1,25-dihydroxyvitamin-D3, and polyamines, as well as methods and parameters for evaluating the level of differentiation achieved.We would appreciate receiving feedback on the protocols described; this will facilitate the development of consensus protocols, which in turn will allow better comparison of data sets generated by different research groups. This continuing standardization, which might be reported on at international meetings like those of IFATS ( http://www.IFATS.org ), might be of benefit for the whole ASC research community.
authors.affiliation: Department of Orthopaedic Surgery, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Robert Jan
authors.initials: RJ
authors.lastname: Kroeze
authors.forename: Marlene
authors.initials: M
authors.lastname: Knippenberg
authors.forename: Marco N
authors.initials: MN
authors.lastname: Helder
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000880:Anthraquinones / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D055419:Bone Morphogenetic Protein 7 / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D006041:Goats; D006801:Humans; D007150:Immunohistochemistry; D058574:Integrin-Binding Sialoprotein / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D010012:Osteogenesis / Q000187:drug effects; D015676:Osteonectin / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013096:Spermine / Q000494:pharmacology; D013194:Staining and Labeling
chemical_list: D000880:Anthraquinones; D055396:Bone Morphogenetic Protein 2; D055419:Bone Morphogenetic Protein 7; D058574:Integrin-Binding Sialoprotein; D015676:Osteonectin; D013096:Spermine; C010078:alizarin; D000469:Alkaline Phosphatase; D002117:Calcitriol