两种重编程系统诱导人牙源性多潜能干细胞的对比研究  

A comparative study of two reprogramming systems for inducing pluripotent stem cells from human dental origin

作　　者：谭小兵[1] 徐静舒[1] 孙贵虎 宋俊呈 戴青原[2] 

机构地区：[1]云南省第一人民医院/昆明理工大学附属医院口腔内科,昆明650032 [2]昆明医科大学第一附属医院心内科,昆明650032

出　　处：《重庆医学》2017年第1期90-93,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(81360161);云南省教育厅基金资助项目(2015Y153)

摘　　要：目的对比研究两种人牙源性多潜能干细胞重编程体系的特点。方法分别利用STEMCCA慢病毒/饲养层和仙台病毒/基质胶两种重编程系统,将人根尖乳头干细胞重编程为iPS细胞,比较两种方法的诱导效率、诱导工作量、iPS细胞非整倍体核型比例、外源性基因消除难易程度等情况。结果STEMCCA重编程体系需要制备饲养层细胞MEF,重编程效率约为0.1%,后期经Cre-Loxp酶切技术可得到无外源性转录基因的iPS细胞。仙台病毒重编程体系为无饲养层培养,基质胶制备方便、标准统一,重编程效率约为0.7%,明显高于STEMCCA系统(P<0.05),外源性病毒和转录基因经自然传代后逐渐消除。结论与STEMCCA系统相比,仙台病毒/基质胶体系更适于人牙源性iPS细胞的诱导。Objective To comparatively study the features of two reprogramming systems of induced pluripotent stem cells （iPSCs） from human dental origin. Methods Two kinds of reprogramming system,i, e. STEMCCA lentivirus /feed layer and Sendai virus /matrigel were used to induce human stem cells from apical papilla（SCAP） into iPSCs, respectively. The induction efficiencies,workload of generating iPSCs,aneuploidy karyotype ratio,complexities of eliminating exogenous transcription factors and specific markers expression were compared between these two systems. Results The STEMCCA reprogramming system required to prepare the feeder cell MEF. The reprogramming efficiency was 0. 1%. Transcription gene-free iPSCs cells were obtained by the Cre-loxp enzyme digestion technique at the later stage. Sendal virus reprogramming system was feeder-free and the preparation of matrigel was quite simple with unified standard. The reprogramming efficiency was 0. 7%, which was much higher than that of STEMCCA system（P〈0.05）. The exogenous virus and transgenes could be gradually eliminated after several passages of natural subclone. Conclusion The Sendal virus/matrigle reprogramming system is much more applicable for the induction of iPSCs from dental origin than the STEMCCA system.

关 键 词：多潜能干细胞 乳头 干细胞 慢病毒属 人诱导性多潜能干细胞 根尖乳头干细胞 重编程 仙台病毒 

分 类 号：R781[医药卫生—口腔医学]