Human Deciduous Teeth Stem Cells (SHED) Display Neural Crest Signature Characters

Abstract

Human dental tissues are sources of neural crest origin multipotent stem cells whose regenerative potential is a focus of extensive studies. Rational programming of clinical applications requires a more detailed knowledge of the characters inherited from neural crest. Investigation of neural crest cells generated from human pluripotent stem cells provided opportunity for their comparison with the postnatal dental cells. The purpose of this study was to investigate the role of the culture conditions in the expression by dental cells of neural crest characters. The results of the study demonstrate that specific neural crest cells requirements, serum-free, active WNT signaling and inactive SMAD 2/3, are needed for the activity of the neural crest characters in dental cells. Specifically, the decreasing concentration of fetal bovine serum (FBS) from regularly used for dental cells 10% to 2% and below, or using serum-free medium, led to emergence of a subset of epithelial-like cells expressing the two key neural crest markers, p75 and HNK-1. Further, the serum-free medium supplemented with neural crest signaling requirements (WNT inducer BIO and TGF-β inhibitor REPSOX), induced epithelial-like phenotype, upregulated the p75, Sox10 and E-Cadherin and downregulated the mesenchymal genes (SNAIL1, ZEB1, TWIST). An expansion medium containing 2% FBS allowed to obtain an epithelial/mesenchymal SHED population showing high proliferation, clonogenic, multi-lineage differentiation capacities. Future experiments will be required to determine the effects of these features on regenerative potential of this novel SHED population.

Establishment and neural differentiation of neural crest-derived stem cells from human dental pulp in serum-free conditions

“…Our findings led us to hypothesize that heterogeneity in our hDPC cultures could lead to inefficacy in differentiation and that an intermediate population may be required for neural maturation, as shown by observations from the literature on hDPC-neural differentiation. 14,[29][30][31] Together with increasing reports obtaining NCSCs from other dental-derived stem cells, 7,8,32,33 it is suggested that a neural crest or neurogenic population needs to be induced or enriched. In this regard, sphere aggregation has been used to enrich for a neural crest-like phenotype in other tissues.…”

“…Nevertheless, most reports are limited to describing the molecular resemblance to neural crest cells, while their actual differentiation capacity is usually left untested, masking their actual translational potential for regenerative medicine. [7][8][9]32,33,37,44 Additionally, it would be important to evaluate a combination of strategies to facilitate the application of oral-derived NCSCs in regenerative medicine, such as their incorporation in advanced manufactured scaffolds or conduits for nerve repair and tissue engineering (eg, bone regeneration). 9,45,46 Finally, the field of dental pulp stem cells and their applications would benefit from addressing specific diseases or targeted applications from preclinically relevant established cultures if we aim to advance the use of dental pulp cells in regenerative medicine.…”

“…Most dental-related stem cell sources are believed to derive from the neural crest, 6 and because of this there is an increasing interest in the study of their neural crest-derived stem cell (NCSC) characteristics. [7][8][9] In contrast to MSCs from non-neural crest origin, these NCSCs could prove particularly useful to restore neural cell types due to their molecular resemblance to embryonic neural crest cells. 10,11 In this regard, there is little information about the identification and isolation of a human NCSC population specifically from human dental pulp cells (hDPCs) of adult, permanent teeth, and only a small number of key direct applications have been reported.…”

SHED-Derived Exosomes Regulate Microglial Polarization in the Treatment of Traumatic Brain Injury in Rats via miR-330-5p

“…In addition to their potential clinical signi cance, our ndings suggest that we may consider the perspective of an embryonic origin of stem cells in the treatment of disease. SHED and other odontogenic stem cells, such as dental pulp stem cells (DPSCs), all originate from neural crest cells during the embryonic stage [24,25]. This may be the reason that the signals inside had better functionality.…”

Prospects of neural crest‐derived cells from oral and dentofacial tissues for application in regenerative medicine

“…A subpopulation of human dental pulp stem cells (hDPSCs) were enriched for the expression of STRO‐1, c‐Kit, and CD34 45 . Furthermore, human deciduous teeth stem cells upregulated p75 and HNK‐1 genes when cultured under low‐serum (<2% FBS) and serum‐free conditions, 46 while 3D culture conditions proved to be effective for maintaining the expression of NC markers, such as nestin, CD271 (p75), and SOX10 47 . Dental pulp contains label‐retaining sphere‐forming cells, which were shown to express specific markers including LNGFR (p75) and nestin 48 .…”