We investigated porcine dental follicle cells at the early crown-formation stage and examined the behavior of cells grown in a collagen type I (Col-I) matrix. Clone-porcine dental follicle cells (DFC-I) and controls, viz., dental follicle itself, nonclone-dental follicle cells, periodontal ligament cells (PDLC), and bone marrow stromal cells, were obtained from 6-month-old pigs. DFC-I showed a different gene expression pattern from controls by reverse-transcription polymerase chain reaction analysis. In addition, Col-I treatment enhanced DFC-I proliferation and increased their alkaline phosphatase activity compared with nontreated DFC-I. The expression of periostin, biglycan, and osteocalcin (OCN) in cells growing on collagen was upregulated, similar to the pattern seen in PDLC. DFC-I with and without Col-I treatment were combined with beta-tricalcium phosphate particles and implanted into immunodeficient mice. Significant differences were found in the gene expression patterns of bone sialoprotein, OCN, and periostin in both treated and non-treated implants at 2 and/or 4 weeks. The results showed that Col-I induced the mineralization pathway in these cells. Hard tissue formation was observed in both implant types at 8 weeks. Our results suggest that Col-I facilitates the differentiation of DFC-I along the mineralization process.