Abstract
Bone marrow-derived mesenchymal stromal stem cells (BMSCs) are a promising cell source for treating articular cartilage defects (Bornes et al., 2014). BMSCs can be seeded within porous biomaterial scaffolds that support three-dimensional cell organization, chondrogenic differentiation and extracellular matrix deposition for the creation of engineered cartilage. This protocol describes our defined methods for isolation and expansion of human and ovine BMSCs, seeding of BMSCs within porous scaffolds and in vitro chondrogenic differentiation (Adesida et al., 2012; Bornes et al., 2015).
Keywords: Mesenchymal stem cell, Scaffold, Cell seeding, Chondrogenesis, Cartilage
Materials and Reagents
Equipment
Procedure
Methods involving manipulation of bone marrow aspirate, cells and porous scaffolds must be performed within a biosafety cabinet. Instruments, solutions and media in contact with bone marrow aspirate, cells and porous scaffolds must be sterile. Solutions and medium should be preheated to 37 °C in a closed container submersed in a water bath.
Recipes
Acknowledgments
Research related to this protocol was funded by the University Hospital Foundation at the University of Alberta Hospital, Canadian Institutes of Health Research and the Edmonton Orthopaedic Research Committee. Integra LifeSciences Corp. and Anika Therapeutics Inc. have generously provided in-kind biomaterials for our laboratory. Stipend support for Troy Bornes was provided by Alberta Innovates - Health Solutions and Canadian Institutes of Health Research.
References
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