牙髓干细胞诱导为神经干细胞方法的初步研究  

A preliminary study on a protocol for neural stem cells from human dental pulp stem cells

作　　者：张欣然 李昊天 黎淼[1] 朱彪 杨文文 唐路[1] ZHANG Xin-ran;LI Hao-tian;LI Miao;ZHU Biao;YANG Wen-wen;TANG Lu(Department of Stomatology,Xuanwu Hospital,Capital Medical University,Beijing 100053,China;Department of Orthopedics,Xuanwu Hospital,Capital Medical University,Beijing 100053,China;Department of Stomatology,Fuxing Hospital,Capital Medical University,Beijing 100045,China)

机构地区：[1]首都医科大学宣武医院口腔科,北京100053 [2]首都医科大学宣武医院骨科,北京100053 [3]首都医科大学附属复兴医院口腔科,北京100045

出　　处：《临床口腔医学杂志》2022年第11期643-647,共5页Journal of Clinical Stomatology

基　　金：国家自然科学基金项目(62005185,62105226);宣武医院国自然青年培育项目(QNPY2020025)。

摘　　要：目的:体外探究牙髓干细胞(dental pulp stem cells,DPSCs)向神经干细胞(neural stem cells,NSCs)诱导分化的有效方式。方法:酶解法分离及培养DPSCs,流式细胞术对DPSCs进行表型分析鉴定。将DPSCs接种于NSCs诱导培养基及低吸附细胞培养板中,每日观察并统计神经球的形态以及直径,对不同直径的神经球进行免疫荧光染色;Western blot检测不同代数DPSCs诱导后Nestin表达情况。结果:在NSCs诱导培养基及低吸附培养板作用下,DPSCs可形成神经球。对神经球形态及直径统计结果表明第5天为最佳诱导时间,6~7 d后神经球形态及直径大小无明显变化。诱导后不同直径大小的神经球Nestin均呈阳性表达,但细胞传代数会影响诱导后神经球Nestin表达。结论:DPSCs可有效的诱导形成NSCs,且早期代数DPSCs诱导后神经球干性较佳。Objective:To investigate the optimal protocol to induce human dental pulp stem cells(DPSCs)into neural stem cells(NSCs).Methods:DPSCs were isolated and cultured through enzymatic digestion.Then the phenotype of DPSCs was analyzed by flow cytometry.DPSCs were induced through NSCs induction medium and low-attachment cell culture plate to neurosphere-like cells.Then the morphology and diameter of neurosphere-like cells were oobserved and counted.The expressions of Nestin in neurosphere with different diameters were detected by immunocytochemistry.The expressions of Nestin in neurosphere induced from DPSCs with differenta passages were detected by Western blot.Results:DPSCs could be induced into neuroshpere-like cells with NSCs induction medium and low-attachment cell culture plate.The best induction time was the 5 th day.Nestin was positively expressed in neurospheres with different diameters after induction,but the passage of DPSCs would affect the expression of Nestin in neurospheres after induction.Conclusion:DPSCs could be induced into NSCs successfully,and the stemness of inducing nerosphere were bettter in the early passage of DPSCs.

关 键 词：牙髓干细胞 神经向诱导 神经球 

分 类 号：R780.2[医药卫生—口腔医学] Q813.1[医药卫生—临床医学]