Development of optimized differentiation media for murine bone marrow multipotent mesenchymal stromal cells

Abstract

Here, we aimed to define adipogenic, osteogenic, and chondrogenic media for the differentiation of murine mesenchymal stromal cells (MSCs). We used differentiation media formulations optimized for use with human bone marrow-derived MSCs as the starting medium and modified the contents of the differentiation media by testing the differentiation potential of the Op9 stromal cell line, a murine enhanced green fluorescent protein transgenic (eGFP+) bone marrow-derived MSC line, and freshly isolated murine BALB/c bone marrow MSCs. We optimized the media through assessment of (immuno)histological staining and assessment of adipogenic, osteogenic, and chondrogenic differentiation-specific gene expression. Adipogenic differentiation was found to improve in a high-glucose setting, whereas chondrogenic differentiation increased in the presence of insulin-like growth factor-1 (IGF-1). Modifications made to the osteogenic media did not further improve the differentiation capacity of the murine MSCs. In conclusion, using minor changes to existing protocols, we found it possible to increase the overall efficacy of murine MSC differentiation. These modified protocols will further aid in a better understanding of the true differentiation capacity of these cells, and improvement of tissue engineering protocols.

Keywords: Differentiation; Multipotent mesenchymal stromal cells; Murine bone marrow; Protocols.