Method: Cells were isolated from deciduous tooth pulp and were grown in DMEM supplemented with 10 % FBS. Cells between 3-5 passages, grown in serum-free medium were characterized for a panel of stem cell markers by RT-PCR. CD117+ cells were magnetically separated and subjected to pancreatic differentiation protocol. Expression of a panel of pancreatically related transcription factors and WNT/ beta catenin signaling pathway were tested with RT-PCR. Immunocytochemistry and flow cytometry of pancreatically related marker were carried out after the differentiation. Concentration of secreted insulin and c-peptide in the medium was determined with ELISA. During the differentiation the cultures were exposed to 0.1ng/mL air of hydrogen sulfide. The cells non-exposed to hydrogen sulfide were used as control.
Result: Insulin, glucagon, somatostatin, and pancreatic polypeptide were positive by immunofluorescence after differentiation. Hydrogen sulfide increased the expression of insulin. WNT signaling pathways were activated by H2S. The concentration of insulin and c-peptide in the medium in H2S sample increased compared with control.
Conclusion: Our results showed that CD117+ fraction of deciduous tooth pulp stem cells are capable of differentiate toward pancreatic endocrine and exocrine cells, and the differentiation of some kinds of pancreatic cells was increased by hydrogen sulfide. Low concentration of hydrogen sulfide may enhance the differentiation property. Human dental pulp cells may therefore have great potential for future cell therapy of pancreatic disorders.