title: NOTCH-Mediated Maintenance and Expansion of Human Bone Marrow Stromal/Stem Cells: A Technology Designed for Orthopedic Regenerative Medicine.
abstract: Human bone marrow-derived stromal/stem cells (BMSCs) have great therapeutic potential for treating skeletal disease and facilitating skeletal repair, although maintaining their multipotency and expanding these cells ex vivo have proven difficult. Because most stem cell-based applications to skeletal regeneration and repair in the clinic would require large numbers of functional BMSCs, recent research has focused on methods for the appropriate selection, expansion, and maintenance of BMSC populations during long-term culture. We describe here a novel biological method that entails selection of human BMSCs based on NOTCH2 expression and activation of the NOTCH signaling pathway in cultured BMSCs via a tissue culture plate coated with recombinant human JAGGED1 (JAG1) ligand. We demonstrate that transient JAG1-mediated NOTCH signaling promotes human BMSC maintenance and expansion while increasing their skeletogenic differentiation capacity, both ex vivo and in vivo. This study is the first of its kind to describe a NOTCH-mediated methodology for the maintenance and expansion of human BMSCs and will serve as a platform for future clinical or translational studies aimed at skeletal regeneration and repair.
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Teng
authors.initials: T
authors.lastname: Long
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Cuicui
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Mirando
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Jianquan
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Regis J
authors.initials: RJ
authors.lastname: O'Keefe
authors.affiliation: Department of Orthopaedics and Rehabilitation, Center for Musculoskeletal Research, and Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York, USA; Department of Orthopaedic Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana, USA; Department of Orthopaedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China; Department of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental, and Genome Laboratories, Duke University School of Medicine, Durham, North Carolina, USA matthew.hilton@dm.duke.edu.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Hilton
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D009985:Orthopedics; D051883:Receptor, Notch2 / Q000096:biosynthesis*; D044968:Regenerative Medicine; D000072099:Serrate-Jagged Proteins; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; C497764:NOTCH2 protein, human; D051883:Receptor, Notch2; D000072099:Serrate-Jagged Proteins
keywords: Bone; Bone marrow stromal cell; Cell culture; Differentiation; Mesenchymal stem cell; NOTCH; Proliferation; Skeleton