In vitro differentiation of human tooth germ stem cells into endothelial‐ and epithelial‐like cells

Abstract

Current clinical techniques in dental practice include stem cell and tissue engineering applications. Dental stem cells are promising primary cell source for mainly tooth tissue engineering. Interaction of mesenchymal stem cell with epithelial and endothelial cells is strictly required for an intact tooth morphogenesis. Therefore, it is important to investigate whether human tooth germ stem cells (hTGSCs) derived from wisdom tooth are suitable for endothelial and epithelial cell transformation in dental tissue regeneration approaches. Differentiation into endothelial and epithelial cell lineages were mimicked under defined conditions, confirmed by real time PCR, western blotting and immunocytochemical analysis by qualitative and quantitative methods. HUVECs and HaCaT cells were used as positive controls for the endothelial and epithelial differentiation assays, respectively. Immunocytochemical and western blotting analysis revealed that terminally differentiated cells expressed cell-lineage markers including CD31, VEGFR2, VE-Cadherin, vWF (endothelial cell markers), and cytokeratin (CK)-17, CK-19, EpCaM, vimentin (epithelial cell markers) in significant levels with respect to undifferentiated control cells. Moreover, high expression levels of VEGFR1, VEGFR2, VEGF, CK-18, and CK-19 genes were detected in differentiated endothelial and epithelial-like cells. Endothelial-like cells derived from hTGSCs were cultured on Matrigel, tube-like structure formations were followed as an indication for functional endothelial differentiation. hTGSCs successfully differentiate into various cell types with a broad range of functional abilities using an in vitro approach. These findings suggest that hTGSCs may serve a potential stem cell source for tissue engineering and cell therapy of epithelial and endothelial tissue.

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering

“…An interesting phenomenon was that BMSCs showed slight CK10 expression in vitro, but this expression was initially negative in vivo and became positive at 30 d. This finding may be explained as an adaptive process of the cells, which could partially lose their original orientation in vivo and, thus, minimise their expression profile until the epithelium became fully differentiated. These results, related to the in vitro differentiation potential of WJSCs and BMSCs, were in agreement with previous work demonstrating their keratinocytic differentiation capability (Garzon et al, 2013) and with the fact that ADSCs and DPSCs have not yet been differentiated into skin keratinocytes -although reports have suggested that these cell types may have epithelial differentiation capability in the presence of inductive medium (Baer et al, 2013;Dogan et al, 2015). In the present study, both WJSCs and BMSCs were able to develop a larger number of cell layers at the in vitro stage, which coincided with the early cytokeratin expression.…”

“…In addition to adipocyte, chondrocyte and osteoblast differentiation potential, MSCs have epithelial differentiation capability and heterotypical models of artificial tissues could be generated using MSCs as alternative cell sources (Garzon et al, 2013). In this regard, preliminary reports support the epithelial differentiation capability of ADSCs (Baer et al, 2013) and human tooth-derived stem cells (Dogan et al, 2015). In turn, WJSCs are efficiently differentiated into cornea epithelial cells (Garzon et al, 2014) and human skin as well as oral mucosa keratinocytes (Garzon et al, 2013).…”

Endothelial committed oral stem cells as modelling in the relationship between periodontal and cardiovascular disease

“…Recently, the angiogenic potential of stem cells derived from dental pulp, gingival fibroblasts, and tooth germ has been demonstrated, putting in evidence that the oral neural crest derived stem cells may be appropriate for generating endothelial tissues (Dogan, Demirci, & Sahin, ; Iohara et al, ; Szepesi et al, ). In our study, we have induced xeno‐free hPDLSCs to endothelial commitment and their differentiation ability was appraised trough CD31 expression evaluated using multiparametric analyses.…”

Dose-dependent Effect of Boric Acid on Myogenic Differentiation of Human Adipose-derived Stem Cells (hADSCs)

“…The flow cytometry analysis of cells was completed using a Becton Dickinson FACS Calibur (Becton Dickinson, San Jose, CA) flow cytometry system. Characterized cells were then differentiated into adipo-, chondro-, and osteogenic lineages to confirm mesenchymal stem cell (MSC) properties as described previously [16]. Briefly, cells were seeded on six well plates at a cell density of 5×10 4 cells/well.…”