The Promise of Human Induced Pluripotent Stem Cells in Dental Research

Induced pluripotent stem cell-based therapy for treating genetic disorders has become an interesting field of research in recent years. However, there is a paucity of information regarding the applicability of induced pluripotent stem cells in dental research. Recent advances in the use of induced pluripotent stem cells have the potential for developing disease-specific iPSC lines in vitro from patients. Indeed, this has provided a perfect cell source for disease modeling and a better understanding of genetic aberrations, pathogenicity, and drug screening. In this paper, we will summarize the recent progress of the disease-specific iPSC development for various human diseases and try to evaluate the possibility of application of iPS technology in dentistry, including its capacity for reprogramming some genetic orodental diseases. In addition to the easy availability and suitability of dental stem cells, the approach of generating patient-specific pluripotent stem cells will undoubtedly benefit patients suffering from orodental disorders

Amenable epigenetic traits of dental pulp stem cells underlie high capability of xeno-free episomal reprogramming

Abstract Background While a shift towards non-viral and animal component-free methods of generating induced pluripotent stem (iPS) cells is preferred for safer clinical applications, there is still a shortage of reliable cell sources and protocols for efficient reprogramming. Methods Here, we show a robust episomal and xeno-free reprogramming strategy for human iPS generation from dental pulp stem cells (DPSCs) which renders good efficiency (0.19%) over a short time frame (13–18 days). Results The robustness of DPSCs as starting cells for iPS induction is found due to their exceptional inherent stemness properties, developmental origin from neural crest cells, specification for tissue commitment, and differentiation capability. To investigate the epigenetic basis for the high reprogramming efficiency of DPSCs, we performed genome-wide DNA methylation analysis and found that the epigenetic signature of DPSCs associated with pluripotent, developmental, and ecto-mesenchymal genes is relatively close to that of iPS and embryonic stem (ES) cells. Among these genes, it is found that overexpression of PAX9 and knockdown of HERV-FRD improved the efficiencies of iPS generation. Conclusion In conclusion, our study provides underlying epigenetic mechanisms that establish a robust platform for efficient generation of iPS cells from DPSCs, facilitating industrial and clinical use of iPS technology for therapeutic needs

Additional file 1: of Amenable epigenetic traits of dental pulp stem cells underlie high capability of xeno-free episomal reprogramming

Table S1. List of commercial and patient-derived dental stem cell samples used. Table S2. DiPS colony counts of dental pulp stem cell samples used for reprogramming under different culture conditions. Figure S1. Transduction efficiencies for reprogramming DPSCs. Figure S2. Generation of iPS cells from DPSCs in the presence of enhancer compounds. Figure S3. Time course during episomal-based reprogramming of DPSCs. Figure S4. Characterisation of DPSCs cultured in xeno-free media. Figure S5. Cytogenetics and in vivo characterisation of DiPS lines generated under xeno-free culture conditions. Table S3. List of differentially methylated regions of selected genes in DPSCs versus ASCs with respect to iPS (DiPS and AiPS) and H1 hES cell lines. Table S4. DNA methylation raw data analysed with multiple probe sets of PAX9 gene that exhibit significant differences between DPSCs and ASCs. Table S5. Top networks by ingenuity pathway analysis (IPA) for differentially methylated genes in ASCs versus AiPS cells that do not exhibit such differences in DPSCs versus DiPS cells. Figure S6. Pluripotent and self-renewal supporting characteristics of DPSCs. (ZIP 8876 kb

Novel live cell fluorescent probe for human-induced pluripotent stem cells highlights early reprogramming population

Background Despite recent rapid progress in method development and biological understanding of induced pluripotent stem (iPS) cells, there has been a relative shortage of tools that monitor the early reprogramming process into human iPS cells. Methods We screened the in-house built fluorescent library compounds that specifically bind human iPS cells. After tertiary screening, the selected probe was analyzed for its ability to detect reprogramming cells in the time-dependent manner using high-content imaging analysis. The probe was compared with conventional dyes in different reprogramming methods, cell types, and cell culture conditions. Cell sorting was performed with the fluorescent probe to analyze the early reprogramming cells for their pluripotent characteristics and genome-wide gene expression signatures by RNA-seq. Finally, the candidate reprogramming factor identified was investigated for its ability to modulate reprogramming efficiency. Results We identified a novel BODIPY-derived fluorescent probe, BDL-E5, which detects live human iPS cells at the early reprogramming stage. BDL-E5 can recognize authentic reprogramming cells around 7 days before iPS colonies are formed and stained positive with conventional pluripotent markers. Cell sorting of reprogrammed cells with BDL-E5 allowed generation of an increased number and higher quality of iPS cells. RNA sequencing analysis of BDL-E5-positive versus negative cells revealed early reprogramming patterns of gene expression, which notably included CREB1. Reprogramming efficiency was significantly increased by overexpression of CREB1 and decreased by knockdown of CREB1. Conclusion Collectively, BDL-E5 offers a valuable tool for delineating the early reprogramming pathway and clinically applicable commercial production of human iPS cells.11Nsciescopu