两种人牙源性iPSCs的重编程效应及生物学特性比较

摘要

目的 比较两种人牙源性iPSCs的重编程效应及其生物学特性.方法 原代培养人牙髓干细胞(DPSCs)和根尖乳头干细胞(SCAP),仙台病毒重编程系统将两种细胞诱导为诱导性多潜能干细胞(iPSCs),比较两种iPSCs的形态、重编程效率及时间,逆转录-聚合酶链反应(RT-PCR)检测SeV及外源性转录基因的表达.结果 人DPSCs、SCAP均重编程为iPSCs,具有典型ES样克隆形态,重编程效率分别为(0.68 ± 0.02)%、(0.7 ± 0.01)%,差异无统计学意义(P> 0.05);重编程时间分别为(26.0 ± 2.1)、(27.0 ± 1.4)d,差异无统计学意义(P>0.05).RT-PCR结果显示,两种iPSCs无外源性病毒或转录基因序列的表达.结论 人DPSCs和SCAP可重编程为无病毒、无转录基因iPSCs,是iPSCs的理想来源.%Objective To compare the reprogramming effects and biological characteristics of two types of human odontogen-ic induced pluripotent stem cells(iPSCs).Methods Human dental pulp stem cells(DPSCs)and stem cells from apical papilla (SCAP)were isolated and primarily cultured.The Sendai reprogramming system was utilized to induce DPSCs and SCAP into iP-SCs.The morphology,reprogramming efficiency,reprogramming and time were compared between human DPSCs-iPSCs and SCAP-iPSCs.The SeV and exogenous transcriptional gene expression were detected by RT-PCR.Results Human DPSCs and SCAP were reprogrammed as iPSCs with classical ES-like clonal morphology.The reprogramming efficiencies were(0.68 ± 0.02)% and(0.7 ± 0.01)% respectively,the difference was not statistically significant(P>0.05).The reprogramming time was(26.0 ± 2.1)d and (27.0 ± 1.4)d respectively,the difference was not statistically significant(P>0.05).The RT-PCR results showed that no expres-sion of exogenous virus or transcriptional gene sequence in both iPSCs.Conclusion Human DPSCs and SCAP can be reprogrammed as virus-free and transgene-free iPSCs,which are the ideal sources of iPSCs.