不同诱导环境对牙周膜干细胞膜片生物学特性的影响  

Effects of different induction systems on periodontal ligament stem cell sheets

作　　者：刘一涵[1] 赵喜聪[2] 张勇杰[2] 刘洪臣[1] 朱斌[2] 

机构地区：[1]中国人民解放军总医院口腔医学研究所,北京100853 [2]第四军医大学口腔医学院

出　　处：《实用口腔医学杂志》2012年第3期279-284,共6页Journal of Practical Stomatology

摘　　要：目的:探讨不同诱导体系对牙周膜干细胞(periodontal ligament stem cells,PDLSCs)膜片的构建以及生物学性能的影响。方法:分离培养大鼠发育期根端复合体(developing apical complex,DAC)细胞,隔天更换收集细胞培养液上清并过滤冻存(DAC条件培养液)。分离培养人PDLSCs,并扩增构建PDLSCs细胞膜片,利用DAC条件培养液和成骨诱导液分别诱导PDLSCs膜片5、10、15 d。通过倒置显微镜、HE染色、免疫组化、RT-PCR检测经过诱导的人PDLSCs细胞膜片形态学和功能学变化。结果:与成骨诱导相比,DAC诱导后膜片含有丰富短棒状细胞和大量细胞外基质(extracellular matrix,ECM),细胞膜片ALP和periostin表达丰富,体外成骨基因表达更高。结论:与成骨诱导体系相比,DAC诱导体系更适合PDLSCs膜片的形成。Objective： To study the effects of different induction systems on the construction and biofunction of periodontal ligament stem cell sheets.Methods： Developing apical complex（DAC） of 14 day-aged SD rats was separated gently and in vitro cultured with α-MEM and 10% FBS.The medium was changed every two days and collected,the supernatant of the used medium was prepared for DAC conditioned induction.Periodontal ligament stem cells（PDLSCs） were obtained and cultured.After identification,PDLSCs were subcuhured,induced by DAC conditioned medium and osteogenectic induction respectively;PDLSC sheets were constructed.Cell sheets were examined by inverted microscope,HE staining,RT-PCR,and immunohistochemistry.Results： PDLSC sheets formed in different induction systems finally.Cell sheet under DAC induction showed more extracellular matrix（ECM） and cells,expressed higher livel of ALP,periostin and osteogenic genes than that under osteogenic induction.Conclusion： DAC induction may promote the formation and biofunction of PDLSC sheet.

关 键 词：牙周膜干细胞 细胞膜片 发育期根端复合体 诱导 

分 类 号：R780.2[医药卫生—口腔医学]