In Vitro Cultivation of Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth in Low-Xenogeneic-Serum Containing Media

Abstract

Introduction and aims: Recently, the human regenerative medicine has been departing from the use of xenogeneic materials for the possibility of zoonosis transfer and development of prion infection. We have focused our research on cultivation of Dental Pulp Stem Cells from Human Exfoliated Deciduous Teeth (SHED) in a medium with lowered concentration of Fetal Calf Serum (FCS), which decreases the chances of the above mentioned negative impacts on the cultivated cell population. In 2011, Karbanova et al. have shown that the Dental Pulp Stem Cells (DPSCs) from Impacted Third Molars can be viably cultured in a xenogeneic medium containing as little as 2% FCS. Our hypothesis, that SHED should react similarly to isolation and cultivation methods as DPSCs, follows from their similarities in the place of origin and their niches. Method: We have tested this hypothesis on three lineages of SHED from varying donours. The SHED were isolated from the exfoliated deciduous teeth dental pulp using enzymatic dissociation and seeded onto cultivation dishes containing the test medium of 2% of FCS. The cells were then continuously expanded over the Hayflick Limit, the proof of "stemness". Following the proliferation potential test, we have examined the phenotype of the cultured undifferentiated cells which expressed high positivity of the Mesenchymal Stem Cell (MSC) surface markers, which agrees with the characteristic SHED phenotype. After we have audited the impact of the isolation and cultivation methods, we have proceeded to test their differentiation potential by seeding the cultured SHED into various differentiation media. The SHED have successfully differentiated into all the targeted unipotent tissue and cell types, namely: osteoblasts, chondrocytes, endothelocytes, myofibroblasts and neural cells. Results: Overall, our data shows that the SHED cultured in low-xenogeneic-serum medium, unlike the SHED cultured in high-xenogeneic-serum media, express lowered positivity of the same surface markers. Conclusion: Our results suggest that SHED are exceptionally promising and available mesenchymal stem cell population, especially applicable for neuroregenerative medicine.

Keywords

• stem cells from human exfoliated deciduous teeth
• shed
• xenogeneic serum
• fetal calf serum
• 2% fcs
• enzymatic digestion
• differentiation potential