title: Expansion on extracellular matrix deposited by human bone marrow stromal cells facilitates stem cell proliferation and tissue-specific lineage potential.
abstract: Our objective was to assess the rejuvenation effect of extracellular matrix (ECM) deposited by human bone marrow stromal cells (hBMSCs) on hBMSC expansion and tissue-specific lineage differentiation potential. Passage 5 hBMSCs were expanded on ECM or conventional plastic flasks (Plastic) for one passage. Cell number was counted and immunophenotype profiles were assessed using flow cytometry. Selected integrins and proliferation-related pathway signals were assessed using Western blot. The expanded cells were evaluated for their chondrogenic potential in a pellet culture system with TGF-β3-containing chondrogenic medium using gross morphology, histology, immunostaining, biochemical analysis, real-time polymerase chain reaction, Western blot, and biomechanical testing. ECM-expanded hBMSCs were further evaluated for their osteogenic potential using Alizarin Red S staining and alkaline phosphatase activity assay and for their adipogenic potential using Oil Red O staining. ECM-expanded hBMSCs exhibited an enhanced proliferation capacity and an acquired robust chondrogenic potential compared to those grown on Plastic. ECM expansion decreased intracellular reactive oxygen species and increased stage-specific embryonic antigen-4 expression in hBMSCs. ECM expansion also upregulated integrins α2 and β5 and induced a sustained activation of Erk1/2 and cyclin D1. Interestingly, upregulation of TGF-β receptor II during cell expansion and chondrogenic induction might be responsible for an enhanced chondrogenic potential in ECM-expanded hBMSCs. We also found that ECM-expanded hBMSCs had an increased osteogenic potential and decreased adipogenic capacity. ECM deposited by hBMSCs may be a promising approach to expand BMSCs from elderly patients for the treatment of large-scale bone defects through endochondral bone formation.
authors: Stem Cell and Tissue Engineering Laboratory, Division of Exercise Physiology, Department of Orthopaedics, West Virginia University, Morgantown, WV 26506-9196, USA. mpei@hsc.wvu.edu
authors: Ming
authors: M
authors: Pei
authors: Fan
authors: F
authors: He
authors: Vincent L
authors: VL
authors: Kish
mesh_terms: D050156:Adipogenesis / Q000235:genetics; D000328:Adult; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D019070:Cell Lineage; D049109:Cell Proliferation; D020219:Chondrogenesis / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D016023:Integrins / Q000378:metabolism; D008297:Male; D009928:Organ Specificity; D010012:Osteogenesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology; D055815:Young Adult
chemical_list: D016023:Integrins