title: Clinical-scale expansion of a mixed population of bone-marrow-derived stem and progenitor cells for potential use in bone-tissue regeneration.
abstract: Preclinical and clinical studies have demonstrated the ability of bone marrow derived stem and progenitor cells to regenerate many tissues, including bone. Methods to expand or enrich progenitors from bone marrow are common; however, these methods include many steps not amenable to clinical use. A closed automated cell production culture system was developed for clinical-scale ex vivo production of bone marrow-derived stem and progenitor cells for hematopoietic reconstitution. The current study tested the ability of this bioreactor system to produce progenitor cells, termed tissue repair cells (TRC), possessing osteogenic potential. Three TRC formulations were evaluated: (a) cells cultured without exogenous cytokines (TRC); (b) cells cultured with exogenous cytokines (TRC-C); and (c) an adherent subset of TRC-C (TRC-C(Ad)). Starting human bone marrow mononuclear cells (BM MNC) and TRC products were characterized for the expression of cell surface markers, in vitro colony forming ability, and in vivo osteogenic potential. Results showed significant expansion of mesenchymal progenitors (CD90+, CD105+, and CD166+) in each TRC formulation. In vivo bone formation, measured by histology, was highest in the TRC group, followed by TRC-C(Ad) and TRC-C. The TRC product outperformed starting BM MNC and had equivalent bone forming potential to purified MSCs at the same cell dose. Post hoc analysis revealed that the presence of CD90+, CD105+, and CD166+ correlated strongly with in vivo bone formation scores (r(2) > .95). These results demonstrate that this bioreactor system can be used to generate, in a single step, a population of progenitor cells with potent osteogenic potential. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: Department of Orthopaedics, Case Western Reserve University, University Hospitals of Cleveland, 6 Floor Hanna Building, 11100 Cedar Avenue, Cleveland, Ohio 44106, USA. James.Dennis@case.edu
authors.forename: James E
authors.initials: JE
authors.lastname: Dennis
authors.forename: Kelly
authors.initials: K
authors.lastname: Esterly
authors.forename: Amad
authors.initials: A
authors.lastname: Awadallah
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Parrish
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Poynter
authors.forename: Kristin L
authors.initials: KL
authors.lastname: Goltry
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D001842:Bone and Bones / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D004921:Erythropoietin / Q000494:pharmacology; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000494:pharmacology; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D016513:Mice, SCID; D008875:Middle Aged; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis / Q000502:physiology*; D019736:Prostheses and Implants; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D007377:Interleukin-3; D008565:Membrane Proteins; C078524:PIXY321 fusion protein, recombinant; D011993:Recombinant Fusion Proteins; C084881:flt3 ligand protein; D004921:Erythropoietin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor