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

Abstract

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 genes 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. © 2015, Semnan University of Medical Sciences. All rights reserved.