title: After repeated division, bone marrow stromal cells express inhibitory factors with osteogenic capabilities, and EphA5 is a primary candidate.
abstract: The differentiation capability of human bone marrow stromal cells (hBMSCs) is thought to deteriorate over multiple doubling processes. To clarify the deterioration mechanisms, the multilineage differentiation capabilities of short- and long-term passaged BMSCs were compared. Predictably, long-term passaged BMSCs showed reduced differentiation capacities compared to short-term passaged cells. Furthermore, a non-human primate heterotopic bone formation model demonstrated that long-term passaged BMSCs have bone formation capabilities but also exert inhibitory effects on bone formation. This finding indicated that long-term passaged BMSCs express higher levels of inhibitory factors than short-term passaged BMSCs do. Co-culture assays of short- and long-term passaged BMSCs suggested that the inhibitory signals required cell-cell contact and would therefore be expressed on the cell membrane. A microarray analysis of BMSCs identified ephrin type-A receptor 5 (EphA5) as an inhibitory factor candidate. Quantitative PCR revealed that among all members of the ephrin and Eph receptor families, only the expression of EphA5 was increased by BMSC proliferation. A gene knockdown analysis using siRNAs demonstrated that knockdown of EphA5 gene expression in long-term passaged BMSCs led to an increase in ALP mRNA expression. These results indicate that EphA5 may be a negative regulator of bone formation. A better understanding of the roles of the ephrin and Eph receptor families in hBMSCs may lead to alternative approaches for manipulating hBMSC fate. In addition, this avenue of discovery may provide new therapeutic targets and quality-control markers of the osteogenic differentiation capabilities of hBMSCs.
authors.affiliation: Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan; Global Center of Excellence (GCOE) Program, International Research Center for Molecular Science in Tooth and Bone Diseases, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Yamada
authors.forename: Masato
authors.initials: M
authors.lastname: Yuasa
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Masaoka
authors.forename: Takashi
authors.initials: T
authors.lastname: Taniyama
authors.forename: Hidetsugu
authors.initials: H
authors.lastname: Maehara
authors.forename: Ichiro
authors.initials: I
authors.lastname: Torigoe
authors.forename: Toshitaka
authors.initials: T
authors.lastname: Yoshii
authors.forename: Kenichi
authors.initials: K
authors.lastname: Shinomiya
authors.forename: Atsushi
authors.initials: A
authors.lastname: Okawa
authors.forename: Shinichi
authors.initials: S
authors.lastname: Sotome
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D008251:Macaca; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D034741:RNA, Small Interfering / Q000378:metabolism; D036123:Receptor, EphA5 / Q000235:genetics / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D017077:Culture Media, Conditioned; D034741:RNA, Small Interfering; D003907:Dexamethasone; C484173:EPHA5 protein, human; D036123:Receptor, EphA5
keywords: Bone marrow stromal cells; Cell–cell contact; EphA5; Long-term culture; Multi-lineage potential; Osteogenesis