As a developmental precursor for diverse periodontal tissues, the dental follicle (DF) harbors great promise for periodontal tissue regeneration. However, development of optimal therapy awaits the answer to a key question that impinges on many issues in development-Do adult progenitor tissues form a homogeneous cell population that differentiates into target tissues when they arrive at the site, or they contain heterogeneous cell populations that are committed to specific fates? To address the homogeneity/heterogeneity question, we analyzed differentiation pathways and markers in several cloned DF cell lines. Our studies revealed that each of our cloned DF lines featured remarkably unique characteristics, indicative of a separate and distinct lineage. One line, DF1, was high in proliferative activity but did not display any mineralization behavior, suggesting that it might be related to a periodontal ligament-type lineage. DF2 was similar to DF1, but featured remarkably high alkaline phosphatase activity indicative of a highly undifferentiated state. DF3 matched the mineralization characteristics of a same stage alveolar bone line AB1 in terms of gene expression and von Kossa staining, indicating that DF3 might be of cementoblastic or alveolar bone osteoblastic lineage. To verify the multilineage potential of the DF for purposes of tissue engineering, a series of differentiation induction experiments was conducted. For identification purposes, characteristics of these heterogeneous follicular progenitor cells were compared with follicle components in tissue sections of the postnatal developing periodontium. The presence of heterogeneous cell populations in the DF mirrors individual developmental pathways in the formation of the dental integument. The profound cellular heterogeneity of the DF as an adult progenitor for tissue regeneration also suggests that heterogeneous cellular constituents might play as much of a role in tissue regeneration as the inducible characteristics of individual lineages might do.