title: In vitro mesenchymal trilineage differentiation and extracellular matrix production by adipose and bone marrow derived adult equine multipotent stromal cells on a collagen scaffold.
abstract: Directed differentiation of adult multipotent stromal cells (MSC) is critical for effective treatment strategies. This study was designed to evaluate the capability of equine MSC from bone marrow (BMSC) and adipose tissue (ASC) on a type I collagen (COLI) scaffold to undergo chondrogenic, osteogenic and adipogenic differentiation and form extracellular matrix (ECM) in vitro. Following determination of surface antigen expression, MSC were loaded into scaffolds in a perfusion bioreactor and loading efficiency was quantified. Cell-scaffold constructs were assessed after loading and 7, 14 and 21 days of culture in stromal or induction medium. Cell number was determined with DNA content, cell viability and spatial uniformity with confocal laser microscopy and cell phenotype and matrix production with light and scanning electron microscopy and mRNA levels. The MSC were positive for CD29 (>90 %), CD44 (>99 %), and CD105 (>60 %). Loading efficiencies were >70 %. The ASC and BMSC cell numbers on scaffolds were affected by culture in induction medium differently. Viable cells remained uniformly distributed in scaffolds for up to 21 days and could be directed to differentiate or to maintain an MSC phenotype. Micro- and ultrastructure showed lineage-specific cell and ECM changes. Lineage-specific mRNA levels differed between ASC and BMSC with induction and changed with time. Based on these results, equine ASC and BMSC differentiate into chondrogenic, osteogenic and adipogenic lineages and form ECM similarly on COLI scaffolds. The collected data supports the potential for equine MSC-COLI constructs to support diverse equine tissue formation for controlled biological studies.
authors.affiliation: Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, Baton Rouge, LA, 70803, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Xie
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Anna
authors.initials: A
authors.lastname: Marsano
authors.forename: Gordana
authors.initials: G
authors.lastname: Vunjak-Novakovic
authors.forename: Yanru
authors.initials: Y
authors.lastname: Zhang
authors.forename: Mandi J
authors.initials: MJ
authors.lastname: Lopez
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D002417:Cattle; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D019070:Cell Lineage / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D003094:Collagen / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D006736:Horses; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D018613:Microscopy, Confocal; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D054457:Tissue Scaffolds
chemical_list: D012333:RNA, Messenger; D003094:Collagen; D004247:DNA
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