Feasibility Study Of Supernumerary Teeth As A Source Of Stem Cells From Apical Papilla

Object To verify the research value of the supernumerary teeth as a source of stem cell from the apical papilla by comparing stem cell from the apical papilla of human supernumerary teeth(SCAP-S)with common dental pulp stem cell from permanent teeth(DPSC)for all the basic features of cell biology and cell differentiation ability differences.To explore the feasibility of using SCAP-S to induce hard tissue formation in vitro by using real-time fluorescence quantitative to compare the expression capacity of osteoblastic related genes between SCAP-S and DPSC,and the changes between before the induction and after 14 days osteoblastic induction of SCAP-S.Method 1.The number of supernumerary teeth available in pediatric dentistry of the oral hospital of Fujian Medical University during 2016-2018 was investigated and statistically analyzed.Analyzing the amount of supernumerary teeth reserve which could separated SCAP in clinical practice.2.Using digestion method SCAP-S and DPSC were isolated and cultured in vitro.3.Detecting the clone formation ability of SCAP-S and DPSC,and the clone formation rate was calculated.Comparing the growth curves of SCAP-S and DPSC after 7 days cultured.4.Using flow cytometry to measure molecular surface markers of SCAP-S so that the source of stem cell type can be analyzed.5.Comparing the ability of multidirectional differentiation between SCAP-S and DPSC by conducting the adipogenic and osteogenic differentiation.6.Using Real-time fluorescence quantitative PCR to compare the differences of the osteogenic related genes expressions between SCAP-S and DPSC,and the changes between before the induction and after 14 days osteoblastic induction of SCAP-S,so as to analyze SCAP-S ability of osteoblastic induction in vitro.Result 1.The samples of supernumerary teeth which were SCAP available were 433 during 3 years research.The patients who needed the extraction of supernumerary teeth usually were young children,most of them were 4 to 10 years old.2.Compared with DPSC,SCAP-S had a significantly faster proliferation rate and stronger activity.3.Both SCAP-S and DPSC cells had the ability of self-cloning.Compared with DPSC,SCAP-S had more clones and stronger ability of self-cloning than DPSC.The 7-day growth curves of the two types of cells were both "S" shaped,which was in line with the growth trend of stem cells,and SCAP-S showed significantly stronger capacity than DPSC.4.SCAP-S belongs to a source of mesenchymal stem cell rather than hematopoietic stem cell.5.Both SCAP-S and DPSC had the ability of multidirectional differentiation,and both had the same ability of lipogenic differentiation,while SCAP-S had significantly stronger osteogenic ability than DPSC.6.Both SCAP-S and DPSC could express osteogenic genes and had good potential for osteogenesis in vitro,with different advantages.SCAP-S showed up-regulated expression of osteoblastic genes before and after osteoblastic induction in vitro which indicated a good ability of osteoblastic induction in vitro.Conclusion SCAP-S is a kind of seed cell which has good potential with a wide source of clinical samples,rapid growth rate,large amount of multiplication,multidirectional differentiation ability,strong ability to induce osteogenesis in vitro,high expression of osteogenic genes,and source of mesenchymal stem cells.SCAP-S has the value of in-depth study.