Collection, Cryopreservation, and Characterization of Human Dental Pulp–Derived Mesenchymal Stem Cells for Banking and Clinical Use

TLDR

Recent studies have shown that mesenchymal stem cells with therapeutic potential can be isolated from extracted dental tissues. The authors investigated the collection, processing, and cryobiological characteristics of MSC from human teeth under current good tissue practices. They isolated viable DPSC from 31 of 40 teeth, and all 8 characterized cultures expressed MSC markers and differentiated into osteogenic, adipogenic, and chondrogenic lineages, while demonstrating that viable cells can be recovered up to 120 h post‑extraction and from cryopreserved intact teeth, indicating that DPSC isolation is feasible for at least five days after extraction and that minimal processing may suffice for clinical banking under cGTP.

Abstract

Recent studies have shown that mesenchymal stem cells (MSC) with the potential for cell-mediated therapies and tissue engineering applications can be isolated from extracted dental tissues. Here, we investigated the collection, processing, and cryobiological characteristics of MSC from human teeth processed under current good tissue practices (cGTP). Viable dental pulp-derived MSC (DPSC) cultures were isolated from 31 of 40 teeth examined. Of eight DPSC cultures examined more thoroughly, all expressed appropriate cell surface markers and underwent osteogenic, adipogenic, and chondrogenic differentiation in appropriate differentiation medium, thus meeting criteria to be called MSC. Viable DPSC were obtained up to 120 h postextraction. Efficient recovery of DPSC from cryopreserved intact teeth and second-passage DPSC cultures was achieved. These studies indicate that DPSC isolation is feasible for at least 5 days after tooth extraction, and imply that processing immediately after extraction may not be required for successful banking of DPSC. Further, the recovery of viable DPSC after cryopreservation of intact teeth suggests that minimal processing may be needed for the banking of samples with no immediate plans for expansion and use. These initial studies will facilitate the development of future cGTP protocols for the clinical banking of MSC.