title: Impact of bladder-derived acellular matrix, growth factors, and extracellular matrix constituents on the survival and multipotency of marrow-derived mesenchymal stem cells.
abstract: We investigate the effect of bladder-derived acellular matrix (ACM) on bone marrow mesenchymal stem cells (BM-MSC) growth, survival, and differentiation, and evaluate the effect of collagen I and IV on BM-MSC differentiation potential to SMC. BM-MSCs isolated from CD1(_) mice were characterized by surface markers and differentiation into different lineages. BM-MSC SMC potential was further evaluated in stem cell medium alone or supplemented with TGF-β1 and recombinant human platelet-derived growth factor (PDGF-BB) on plastic, collagen I and IV using western blot. Furthermore, BM-MSCs were seeded on porcine derived ACM-fortified with hyaluronic acid and cultured in Mesencult+-growth factors, bone, or fat induction media for 3 weeks. Seeded constructs were evaluated by H&E, Ki67 assay, Oil red O, and Alizarin red stain. SMC differentiation was semiquantified via immunohistochemistry. BM-MSCs differentiated into fat and bone when induced. In Mesencult, BM-MSCs differentiated into SMC, expressing α-SMA, calponin, and MHC. BM-MSCs cultured on collagen I and IV reduced expression of SMC and MHC compared to plastic. On ACM-HA, BM-MSCs maintained multipotent state by differentiating to bone and fat when induced. In Mesencult, BM-MSC-seeded ACM-HA expressed α-SMA, calponin, and MHC. TGF-β1 and PDGF-BB enhanced SMC differentiation on collagens and ACM-HA. SMC proteins expression by BM-MSC varies depending on culture substrate. SMC markers are expressed higher on plastic and lower on collagen I, IV, and ACM-HA, suggesting these substrates preferentially maintain undifferentiated state of BM-MSC, which could be advantageous for incorporation of cell-seeded grafts to permit host modulation of tissue regeneration.
authors.affiliation: Developmental and Stem Cell Biology, Research Institute, The Hospital for Sick Children, Toronto, Canada.
authors.forename: Roula
authors.initials: R
authors.lastname: Antoon
authors.forename: Herman
authors.initials: H
authors.lastname: Yeger
authors.forename: Yasir
authors.initials: Y
authors.lastname: Loai
authors.forename: Syed
authors.initials: S
authors.lastname: Islam
authors.forename: Walid A
authors.initials: WA
authors.lastname: Farhat
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D024042:Collagen Type I / Q000494:pharmacology; D024141:Collagen Type IV / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D010641:Phenotype; D010969:Plastics; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology*; D034421:Sus scrofa; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D001743:Urinary Bladder / Q000378:metabolism*; D000096985:Calponins
chemical_list: D000199:Actins; D000077214:Becaplermin; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D024042:Collagen Type I; D024141:Collagen Type IV; D003470:Culture Media; D006820:Hyaluronic Acid; D008840:Microfilament Proteins; D018995:Myosin Heavy Chains; D010969:Plastics; D020574:Proto-Oncogene Proteins c-sis; D053773:Transforming Growth Factor beta1; D000096985:Calponins