naa a22 4500 510739016 CHVBK 20180411083021.0 cr unu---uuuuu 180411e20131201xx s 000 0 eng 10.1007/s12015-013-9456-1 doi (NATIONALLICENCE)springer-10.1007/s12015-013-9456-1 In Vitro Mesenchymal Trilineage Differentiation and Extracellular Matrix Production by Adipose and Bone Marrow Derived Adult Equine Multipotent Stromal Cells on a Collagen Scaffold [Elektronische Daten] [Lin Xie, Nan Zhang, Anna Marsano, Gordana Vunjak-Novakovic, Yanru Zhang, Mandi Lopez] 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 21days 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 21days 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. The Author(s), 2013 Adipose nationallicence Bone marrow nationallicence Multipotent stromal cell nationallicence Equine nationallicence Extra-cellular matrix nationallicence Bioreactor nationallicence Osteogenesis nationallicence Adipogenesis nationallicence Chondrogenesis nationallicence Xie Lin Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut Zhang Nan Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut Marsano Anna Department of Biomedical Engineering, Columbia University, 10032, New York, NY, USA aut Vunjak-Novakovic Gordana Department of Biomedical Engineering, Columbia University, 10032, New York, NY, USA aut Zhang Yanru Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut Lopez Mandi Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/6(2013-12-01), 858-872 1550-8943 9:6<858 2013 9 12015 https://doi.org/10.1007/s12015-013-9456-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12015-013-9456-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xie Lin Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut NATIONALLICENCE 700 1- Zhang Nan Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut NATIONALLICENCE 700 1- Marsano Anna Department of Biomedical Engineering, Columbia University, 10032, New York, NY, USA aut NATIONALLICENCE 700 1- Vunjak-Novakovic Gordana Department of Biomedical Engineering, Columbia University, 10032, New York, NY, USA aut NATIONALLICENCE 700 1- Zhang Yanru Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut NATIONALLICENCE 700 1- Lopez Mandi Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, 70803, Baton Rouge, LA, USA aut NATIONALLICENCE 773 0- Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/6(2013-12-01), 858-872 1550-8943 9:6<858 2013 9 12015 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer