人牙龈成纤维细胞诱导分化为神经样细胞的实验研究  

Experiment of human gingival fibroblasts induction into nerve-like cells

作　　者：苏建芳 宋鹏[1] 董福生[1] 刘慧娟 张艳宁[1] 王洁[1] SU Jianfang;SONG Peng;DONG Fusheng;LIU Huijuan;ZHANG Yanning;WANG Jie(Department of Pathology,School and College of Stomatology,Hebei Medical University,The Key Laboratory of Hebei Province,Hebei Clinical Research Center for Oral Diseases,Shijiazhuang 050017)

机构地区：[1]河北医科大学口腔医学院·口腔医院病理科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,石家庄050017

出　　处：《现代口腔医学杂志》2021年第6期361-366,共6页Journal of Modern Stomatology

基　　金：河北省政府资助临床医学优秀人才培养和基础课题研究项目(361029)。

摘　　要：目的探讨作为结缔组织的人牙龈成纤维细胞,分化为神经样细胞的可能性。方法留取阻生齿拔除后,切取的正常牙龈组织,体外原代培养人牙龈成纤维细胞,采用抗vimentin、GFAP和MAP-2进行细胞鉴定。采用低糖DMEM,含1mmol/Lβ-ME,20μmol/L RA,2%DMSO诱导培养人牙龈成纤维细胞向神经样细胞分化。应用ELISA检测神经样细胞分泌的神经递质。结果人牙龈组织块培养5~7天,细胞爬出。传代后细胞鉴定发现,成纤维细胞呈梭形,vimentin呈阳性,胞质棕黄色;GFAP和MAP-2呈阴性,胞质未着色。诱导培养12天后,细胞胞体变为三角形或多角形,突起2~3个,细而长,突起之间互相连接成网状。神经样细胞GFAP、MAP-2、vimentin均呈阳性表达。ELISA结果显示,神经样细胞具有分泌神经递质乙酰胆碱和多巴胺的功能,与对照组之间差别均有显著性(P<0.05)。结论(1)人牙龈成纤维细胞能够诱导分化为神经样细胞。(2)诱导形成的神经样细胞具有分泌神经递质乙酰胆碱和多巴胺的功能。Objective To investigate the possibility of fibroblasts from human gingiva,to differential into nervelike cells.Methods The normal human gingiva was collected from a patient with an extraction of impacted tooth.The primary culture of human gingival fibroblasts was in low-glucose DMEM containing fetal bovine serum.Immunocytochemical stains against vimentin,GFAP and MAP-2 respectively.The gingival fibroblasts were induced with 1 mmol/Lβ-ME,20μmol/L RA,2%DMSO and induced for 12 days.Enzyme linked immunosorbent assay(ELISA)was used to test the neurotransmitter secreted by the nerve-like cells.Results After cultured for 5~7 days,the human gingival fibroblasts were around the gingival tissue.The elongation cells with round or ellipse nucleus and 2~3 nucleolus grows vortex in shape.Immunochemical stains showed that vimentin was positive strongly in the gingival fibroblasts,and GFAP and MAP-2 were negative in the gingival fibroblasts.After induced for 12 days,most of the fibroblasts were changed into triangular or polygonal in shape with single or several protrusions which were long and thin,separated or connected into a network.The nerve-like cells showed positive for human vimentin,GFAP and MAP-2.Dopamine and acetylcholine could be detected by the nerve-like cells by the ELISA.Furthermore,the concentration of both two kinds of neurotransmitter between the control and nerve-like cells showed statistical differences(P<0.05).Conclusion(1)The human gingival fibroblasts have the capability of being induced into nerve-like cells.(2)The nerve-like cells could produce the neurotransmitter dopamine and acetylcholine.

关 键 词：成纤维细胞 牙龈 神经样细胞 细胞培养 诱导 

分 类 号：R780.2[医药卫生—口腔医学]