In Vitro Mesenchymal Trilineage Differentiation and Extracellular Matrix Production by Adipose and Bone Marrow Derived Adult Equine Multipotent Stromal Cells on a Collagen Scaffold
Open Access
- 28 July 2013
- journal article
- research article
- Published by Springer Science and Business Media LLC in Stem Cell Reviews and Reports
- Vol. 9 (6), 858-872
- https://doi.org/10.1007/s12015-013-9456-1
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.Keywords
This publication has 66 references indexed in Scilit:
- Proliferation as a Requirement for In Vitro Chondrogenesis of Human Mesenchymal Stem CellsStem Cells and Development, 2012
- Improved cartilage repair viain vitropre-maturation of MSC-seeded hyaluronic acid hydrogelsBiomedical Materials, 2012
- SOX2 has a crucial role in the lineage determination and proliferation of mesenchymal stem cells through Dickkopf-1 and c-MYCCell Death & Differentiation, 2011
- Impaired expansion and multipotentiality of adult stromal cells in a rat chronic alcohol abuse modelAlcohol, 2011
- Transglutaminase 2 regulates early chondrogenesis and glycosaminoglycan synthesisMechanisms of Development, 2011
- Evaluating osteochondral defect repair potential of autologous rabbit bone marrow cells on type II collagen scaffoldCytotechnology, 2010
- Stromal stem cells from adipose tissue and bone marrow of age‐matched female donors display distinct immunophenotypic profilesJournal of Cellular Physiology, 2010
- Temporal Analysis of Equine Bone Marrow Aspirate During Establishment of Putative Mesenchymal Progenitor Cell PopulationsStem Cells and Development, 2010
- Perfusion seeding of channeled elastomeric scaffolds with myocytes and endothelial cells for cardiac tissue engineeringBiotechnology Progress, 2010
- Acceleration of spinal fusion using syngeneic and allogeneic adult adipose derived stem cells in a rat modelJournal of Orthopaedic Research, 2009