ISOLATION, GROWTH AND ODONTOGENIC DIFFERENTIATION OF HUMAN DENTAL PULP STEM CELLS IN VITRO

Authors

Nicholas Bauter, University of the Pacific
Justin Hannon, University of the Pacific
Bahram Javid, University of the Pacific
Anders Nattestad, University of the PacificFollow
Abir Balgonaim, University of the Pacific
Michelle Bittner-Eberle, University of the Pacific
Kiri Herchold-Yee, University of the Pacific
Mirek Tolar, University of the Pacific

Lead Author Affiliation

Dugoni School of Dentistry

Second Author Affiliation

Dugoni School of Dentistry

Third Author Affiliation

Dugoni School of Dentistry

Fourth Author Affiliation

Dugoni School of Dentistry

Fifth Author Affiliation

Dugoni School of Dentistry

Sixth Author Affiliation

Dugoni School of Dentistry

Introduction/Abstract

Adult dental pulp-derived stem cells (DPSC’s) can differentiate into mesoderm-derived cell types and nerve cells. DPSC’s are excellent candidates for regenerative cell therapy restorations in the craniofacial region. For this to be possible, the DPSC’s must be efficiently expanded in vitro with no loss of their differentiation potential. We examined effects of combinations of platelet-derived factors (PDF) and fibrin substrate on multiplication and odontogenic differentiation of DPSC’s in vitro. A response of DPSC’s to low oxygen was also tested.

Method

DPSC’s were isolated from vital asymptomatic third molars and were grown on the following substrates: polystyrene, fibrin and fibrin with PDF. The growth medium consisted of basal medium (Lonza) supplemented with human adult serum (10%), L-glutamine and antibiotics. The odontogenic differentiation medium consisted of basal medium (Lonza) supplemented with human adult serum (10%), L-glutamine, dexamethasone, L-ascorbic acid, KH2PO4 and antibiotics. The cells were stained for markers of differentiation and mineralization.

Results

We constructed growth curves showing that the substrate containing both fibrin and PDF shortened the lag phase of growth and increased the multiplication and differentiation rate of the DPSC’s most efficiently. The fibrin substrate was second in its efficiency. Strong staining with alizarin red S for mineralization of extracellular matrix and strong staining of cellular alkaline phosphatase documented a successful differentiation. Hypoxia significantly increased proliferation rate of DPSC’s.

Significance

Fibrin, PDF and hypoxia – factors that are normally present in a healing wound - can be utilized to accelerate preparation of DPSC’s for clinical applications.

Location

DeRosa University Center, Stockton campus, University of the Pacific

Format

Poster Presentation