title: Human adipose-derived mesenchymal stem cells: direction to a phenotype sharing similarities with the disc, gene expression profiling, and coculture with human annulus cells.
abstract: Biologic therapies for disc degeneration hold great promise as an emerging concept. Due to ease of harvest and abundance, adipose derived-mesenchymal stem cells (AD-MSC) are a readily available cell source for such therapies. Our objectives in this study were (1) to develop/validate methods to harvest AD-MSC and direct them to a disc-like phenotype by three-dimensional (3D) culture and transforming growth factor (TGF)-beta3 exposure, (2) to assess cell phenotypes with gene expression profiling for these human AD-MSC and annulus cells, and (3) to test whether disc cell-AD-MSC coculture could augment glycosaminoglycan (GAG) production. When AD-MSC were exposed to TGF-beta3, greater extracellular matrix was formed containing types I and II collagen, keratan sulfate, and decorin. Biochemical GAG measurement showed that production was significantly greater in TGF-beta3-treated AD-MSC in 3D culture versus untreated controls (p < 0.05). Gene expression patterns in AD-MSC were compared to annulus cells; 4424 genes were significantly upregulated, and 2290 genes downregulated. Coculture resulted in a 44% greater GAG content compared with AD-MSC or annulus culture alone (p = 0.04). Data indicated that human AD-MSC can successfully be manipulated in 3D culture to express gene products important in the disc, and that coculture of annulus cells with AD-MSC enhances total GAG production.
authors.affiliation: Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina 28232, USA. helen.gruber@carolinashealthcare.org
authors.forename: Helen E
authors.initials: HE
authors.lastname: Gruber
authors.forename: Ray
authors.initials: R
authors.lastname: Deepe
authors.forename: Gretchen L
authors.initials: GL
authors.lastname: Hoelscher
authors.forename: Jane A
authors.initials: JA
authors.lastname: Ingram
authors.forename: H James
authors.initials: HJ
authors.lastname: Norton
authors.forename: Brian
authors.initials: B
authors.lastname: Scannell
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: Loeffler
authors.forename: Natalia
authors.initials: N
authors.lastname: Zinchenko
authors.forename: Edward N
authors.initials: EN
authors.lastname: Hanley
authors.forename: Hazel
authors.initials: H
authors.lastname: Tapp
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology
chemical_list: D006025:Glycosaminoglycans; D053782:Transforming Growth Factor beta3