Abstract
Dental pulp (DP) can be extracted from child’s primary teeth (deciduous), whose loss occurs spontaneously by about 5 to 12 years. Thus, DP presents an easy accessible source of stem cells without ethical concerns. Substantial quantities of stem cells of an excellent quality and at early (2–5) passages are necessary for clinical use, which currently is a problem for use of adult stem cells. Herein, DPs were cultured generating stem cells at least during six months through multiple mechanical transfers into a new culture dish every 3–4 days. We compared stem cells isolated from the same DP before (early population, EP) and six months after several mechanical transfers (late population, LP). No changes, in both EP and LP, were observed in morphology, expression of stem cells markers (nestin, vimentin, fibronectin, SH2, SH3 and Oct3/4), chondrogenic and myogenic differentiation potential, even after cryopreservation. Six hours after DP extraction and in vitro plating, rare 5-bromo-2′-deoxyuridine (BrdU) positive cells were observed in pulp central part. After 72 hours, BrdU positive cells increased in number and were found in DP periphery, thus originating a multicellular population of stem cells of high purity. Multiple stem cell niches were identified in different zones of DP, because abundant expression of nestin, vimentin and Oct3/4 proteins was observed, while STRO-1 protein localization was restricted to perivascular niche. Our finding is of importance for the future of stem cell therapies, providing scaling-up of stem cells at early passages with minimum risk of losing their “stemness”.
Highlights
Isolation of stem cells (SC) from human adult and deciduous teeth has been reported in the last decade [1,2]
Transmission electron microscopy revealed two types of IDPSCs morphology: embryonic stem (ES)-like cells with low cytoplasm-to-nucleus ratio, low cytoplasm density, which are poor of organelles (Figure 1D)
IDPSCs of mesenchymal stem cells (MSCs)-like cells have a high number of stretched out pseudopodes, which serve to explore substrate and more cytoplasm and organelles when compared with IDPSCs of ES-like cells (Figure 1D, E)
Summary
Isolation of stem cells (SC) from human adult and deciduous teeth has been reported in the last decade [1,2]. It has been demonstrated that the use of different handling methods of dental pulp (DP) can lead to the isolation of SC populations with distinct properties These DTSC populations are similar to mesenchymal stem cells (MSCs) or epithelial SCs or they are composed by a mixed population of both cell types [3]. Along with MSC markers, IDPSCs express embryonic stem (ES) cells markers (Oct3/4, Nanog and Sox2) and undergo spontaneous differentiation into a wide range of cell types in vitro [4] These cells showed expressive capacity to contribute into multiple tissues in response to the cellular milieu during human/mouse pre-termed chimeras development [5]. IDPSCs and other dental stem/progenitor cells were recently used to obtain induced pluripotent SCs [9,10] These cells demonstrated higher efficiency of reprogramming than fibroblasts, providing a model for the study of pediatric diseases and disorders. These data strongly suggest that IDPSCs are a hopeful source for the future of SC therapies [11]
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