Int. J. Dev. Biol. 55: 189 - 195 (2011)

https://doi.org/10.1387/ijdb.103090nn

Vol 55, Issue 2

Induced in vitro differentiation of neural-like cells from human exfoliated deciduous teeth-derived stem cells

Original Article | Published: 8 June 2011

Nosrat Nourbakhsh¹, Mitra Soleimani², Zahra Taghipour², Khadijeh Karbalaie³, Seeid-Behrouz Mousavi⁴, Ardeshir Talebi⁵, Fatemeh Nadali⁵, Somayeh Tanhaei³, Gholam-Abbas Kiyani¹, Marziyeh Nematollahi³, Farzaneh Rabiei¹, Mohammad Mardani², Hamid Bahramiyan², Mahmood Torabinejad⁶, Mohammad-Hossein Nasr-Esfahani³ and Hossein Baharvand*^,7,8

¹Pediatric Department, School of Dentistry, ²Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran, ³Department of Cell and Molecular Biology, Royan Institute for Animal Biotechnology, Isfahan, Iran, ⁴Endodentic Department, School of Dentistry, Torabinejad Dental Center, ⁵Pathology Department, Faculty of Medical Sciences, Isfahan University, Isfahan, Iran, ⁶Endodentic Department, Lumalinda University, U.S.A, ⁷Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran and 8Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran.

Abstract

Stem cells from human exfoliated deciduous teeth (SHED) are highly proliferative, clonogenic and multipotent stem cells with a neural crest cell origin. Additionally, they can be collected with minimal invasiveness in comparison with other sources of mesenchymal stem cells (MSCs). Therefore, SHED could be a desirable option for potential therapeutic applications. In this study, SHEDs were established from enzyme-disaggregated deciduous dental pulp obtained from 6 to 9 year-old children. The cells had typical fibroblastoid morphology and expressed antigens characteristic of MSCs, STRO1, CD146, CD45, CD90, CD106 and CD166, but not the hematopoietic and endothelial markers, CD34 and CD31, as assessed by FACS analysis. Differentiation assessment revealed a strong osteogenic and adipogenic potential of SHEDs. In order to further evaluate the in vitro differentiation potential of SHED into neural cells, a simple short time growth factor-mediated induction was used. Immunofluorescence staining and flow cytometric analysis revealed that SHED rapidly expressed nestin and b-III tubulin, and later expressed intermediate neural markers. In addition, the intensity and percentages of nestin and b-III tubulin and mature neural markers (PSA-NCAM, NeuN, Tau, TH, or GFAP) increased significantly following treatment. Moreover, RT-PCR and Western blot analyses showed that the neural markers were strongly up-regulated after induction. In conclusion, these results provide evidence that SHED can differentiate into neural cells by the expression of a comprehensive set of genes and proteins that define neural-like cells in vitro. SHED cells might be considered as new candidates for the autologous transplantation of a wide variety of neurological diseases and neurotraumatic injuries.

Keywords

dental pulp stem cell, Stem cells from Human Exfoliated Deciduous Teeth (SHED)