Cartilage Induction from Mouse Mesenchymal Stem Cells in High-density Micromass Culture

Mesenchymal stem cells have the ability to differentiate into multiple lineages, including adipocytes, osteoblasts and chondrocytes. Mesenchymal stem cells can be induced to differentiate into chondrocytes in extracellular matrices, such as alginate or collagen gel. Mesenchymal stem cells in a cell pellet or micromass culture can be also induced to form cartilages in a defined medium containing chondrogenic cytokines, such as transforming growth factor-β (TGF-β). Here, we describe a simple method to form cartilage by seeding mesenchymal cells derived from limb-bud cells at high cell density. First, we dissected the limb buds from embryonic mice (embryonic day 12.5) and digested them with enzymes (dispase and collagenase). After filtration using a cell strainer, we seeded the cells at high density. Unlike other methods, the method described here is simple and does not require the use of specialized equipment, expensive materials or complex reagents.