Human Dental Pulp Stem Cells – Isolation and Long Term Cultivation

Abstract

Summary: Human adult mesenchymal stem cells (MSCs) are rare elements living in various organs (e.g. bone marrow, skeletal muscle), with capability to differentiate in various cell types (e.g. chondrocytes, adipocytes and osteoblasts). In the year 2000, Gronthos and co-workers isolated stem cells from the human dental pulp (DPSCs). Later on, stem cells from exfoliated tooth were also obtained. The aims of our study were to establish protocol of DPSCs isolation and to cultivate DPSCs either from adult or exfoliated tooth, and to compare these cells with mesenchymal progenitor cell (MPCs) cultures. MPCs were isolated from the human bone marrow of proximal femur. DPSCs were isolated from deciduous and permanent teeth. Both cell types were cultivated under the same conditions in the media with 2 % of FCS supplemented with PDGF and EGF growth factors. We have cultivated undifferentiated DPSCs for long time, over 60 population doublings in cultivation media designed for bone marrow MPCs. After reaching Hayflick's limit, they still have normal karyotype. Initial doubling time of our cultures was from 12 to 50 hours for first 40 population doublings, after reaching 50 population doublings, doubling time had increased to 60-90 hours. Regression analysis of uncumulated population doublings proved tight dependence of population doublings on passage number and slow decrease of proliferation potential. In comparison with bone marrow MPCs, DPSCs share similar biological characteristics and stem cell properties. The results of our experiments proved that the DPSCs and MPCs are highly proliferative, clonogenic cells that can be expanded beyond Hayflick's limit and remain cytogenetically stable. Moreover we have probably isolated two different populations of DPSCs. These DPSCs lines differed one from another in morphology. Because of their high proliferative and differentiation potential, DPSCs can become more attractive, easily accessible source of adult stem cells for therapeutic purposes.

Study on Apoptosis in Human Deciduous Tooth Pulp Cells

“…Higher cellular proliferation capacity was observed in cells originating from deciduous tooth pulp than in cells originating from permanent tooth pulp, which was consistent with prior reports [9][10][11] .…”

Mechanical Stretch Increases the Proliferation While Inhibiting the Osteogenic Differentiation in Dental Pulp Stem Cells

“…DPSCs showed that high expressions of CD29 and CD90, which were common stem cell markers in mesenchymal stem cells and DPSCs. 27,28 DPSCs also showed relatively high expressions of CD49d, while lacked the expressions of CD34 and CD45. 29 …”

Scaffolds for Peripheral Nerve Regeneration, the Importance of In Vitro and In Vivo Studies for the Development of Cell-Based Therapies and Biomaterials: State of the Art

“…DPSCs can be isolated from both the perivascular and the sub-odontoblastic compartments (the inner surface of the tooth and the outer part of the pulp tissue), by separate digestion of the tooth and extracted pulp tissue. Both populations presented identical cell size, doubling times and karyotype stability, differing only in morphology with rounder cells in the sub-odontoblastic compartment versus spindle-shaped cells with long processes in the perivascular one [239]. Pre-clinical experiments have demonstrated that stem cells show promising results in differentiation into neuronal-like cells [226,228,232,233,240] and their secretion of growth factors [229,241,242].…”