Isolation and identification of mesenchymal and neural crest characteristics of dental pulp derived stem cells

  Introduction: Mesenchymal stem cells (MSC) are considered as a unique source of adult stem cells which are usually accessible with minimum ethical concerns. MSCs can be used in stem cell biology , different to other cells, tissue engineering, regenerative medicine and future of cancer treatment researches. The aim of the present study was to isolate dental pulp derived stem cells (DPSCs) , characterizing them with common techniques and determining neural crest gene s expression profile.   Materials and Methods: After collecting the teeth , the pulp chamber exposed and the pulp tissue digested with collagenase IV. Dental pulp derived cells were resuspended and incubated in alpha modified minimum essential medium eagle ( αMEM ) supplemented with 15 % fetal bovine serum at 37°C temperature, 95% humidity and 5% CO2 . The osteogenic and adipogenic differentiation potentials for dental pulp derived cells were assessed in appropriate conditions. Flow-cytometry analyses were utilized to evaluate the percentage of CD90 and CD45 positive markers within isolated cells . The expression of p75ngfr, slug and sox10 genes was measured via RT-PCR.   Results: Dental pulp derived cells were appropriately attached to the culture vessels and showed high proliferation rate on culture medium . Cells could also differentiate to osteogenic and adipose cells displayed by Alizarin red and Oil red staining . Higher percentage of pulp derived cells were CD90 positive , while the percentage of CD45 expression, as a marker of hematopoietic cells, was negligible . Furthermore , the RT-PCR also revealed the expression of p75ngfr, slug and sox10 in the MSCs isolated from the dental pulps.   Conclusion: Dental pulp derived cell have the mesenchymal stem cells characteristics and at least a portion of them have neural crest origin and identity