Recently, a variety of mesenchymal stem cells (MSCs), including dental pulp stem cells, stem cells from human exfoliated deciduous teeth, stem cells from apical papilla, periodontal ligament stem cells, and mesenchymal stem cells derived from human gingival, were isolated from orofacial and dental tissues. However, it is unknown whether these orofacial stem cells are derived from mesoderm or neural crest cell. In order to encourage orofacial MSC investigation, we provide detailed protocols for assessing lineage -differentiation of orofacial MSCs.