Generation Of Larger Teeth From Human IPS Cells With Pig Dental Mesenchyme

As a new medical strategy,induced pluripotent stem cells(iPS cells)have broad prospects in the field of dental restoration.A variety of tooth derived cell types have been shown to be transformed into iPS cells.Meanwhile,these iPS cells also have tissue memory,making the induction process easier and more effective.Previous experiments successfully induced human iPS cells into epithelial cells which have odontoblast potential.After co culture with the fetal mouse mesenchyme,the recombinant was transplanted into the nude mice and tooth like structures were found 3 weeks after.Accordingly,in order to find out the feasibility of the regeneration model and the effects of the mesenchyme on the tooth regeneration,the further study was been carried out.In this study,induced pluripotent stem cells derived from human urine were differentiated into epithelioid cells.The epithelial cells were collected at 0 to 28 days and detected by qPCR and immunofluorescence for the transplantation time searching.The oral mesenchymal isolated from fetal porcine tooth germ at embryonic day 48 and separated when digested,recombined with epithelial sheet co culture for 1-2 days before transplanted.After that,we compared the related data of regenerated tooth from porcine mesenchymal and mouse mesenchymal.The results showed that the epithelial cell layer induced by human urine derived iPS cells was most capable of binding at seventh days.After the epithelial cell layer was recombined with the fetal porcine oral mesenchyme,the recombinant was transplanted into the renal capsule and the teeth were regenerated 20 weeks after.Statistics showed that tooth forming rate of pig tooth germ group was 92.86%,the tooth forming rate of pig recombinant was 72.97%,much higher than the mouse recombinant group(21.81%).At the same time,all the regenerated dental tissues from pig are larger than the mouse.At the 20 week of the experiment,the epithelial cells differentiated into enamel cells and were associated with dental pulp formation.In conclusion,our experiments show that our iPS tooth regeneration model is well suited and oral mesenchyme has a great effect on tooth regeneration.Has provided a great foundation for our subsequent use of human iPS cells for teeth regeneration,also deepens our understanding of oral mesenchyme and promotes the use of dental regeneration in later experiments.