Isolation and culture of dental papilla cells from pig

Objective: To isolate and culture the dental papilla cells from postnatal porcine tooth buds. Methods: Dental papillae were dissected and digested by collagenase type I and Dispase II. The single-cell suspensions were obtained and cultured in the medium of DMEM/F12 containing 10% fetal bovine serum. The following steps were performed: 1) Observe the morphological characterizations of dental papilla cells(DPCs) by light microscopy and transmission electron microscopy (TEM). 2) Analyze the cell cycle through flow cytometry. 3) Identify the dental papilla cells by immunohistochemistry methods . Results: The dental papilla cells were cultured in vitro through the method of enzyme digestion . The cells showed the characteristics a little like fibroblast and bone marrow stromal cells (BMSCs). Most of the cells were in quiescence based on ratio of G0/G1 phase and the proteins of Vimentin, Col I was positively expressed, Col III was negatively expressed.Conclusion: Porcine dental papilla cells can be isolated effectively through digesting method using collagenase type I and Dispase II and cultured effectively in the medium of DMEM/F12 system. Our further research will focus on reconstruction dentin-pulp complex using DPCs and scaffolds.