犬牙囊干细胞膜片的构建及生物学特性的研究  

Construction of Dental Follicle Stem Cell Sheet and its Biological Characteristics in Beagle Dogs

作　　者：黄闯[1] 宋镜明[1] 宋扬[1] 刘佳[1] 王丽颖[1] 金作林[1] 

机构地区：[1]第四军医大学附属口腔医院正畸科,陕西西安710032

出　　处：《现代生物医学进展》2012年第2期226-230,220,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30973358)

摘　　要：目的:利用犬牙囊干细胞(Dental Follicle Stem Cells,DFSCs)构建细胞膜片并研究其生物学特性。方法:取4至6月龄犬尖牙牙胚,分离培养DFSCs,鉴定。用含抗坏血酸的培养基诱导2周构建细胞膜片,并通过倒置显微镜、HE染色、茜素红染色、油红染色、扫描电镜(SEM)对膜片进行形态学检测,检测成骨、成脂能力。结果:DFSCs于体外被成功分离、纯化、培养,细胞克隆形成率约为5.1%。流式鉴定为CD29+CD44+CD34-,增殖能力及克隆形成能力较强,并能成功构建成细胞膜片。光镜和电镜显示膜片细胞排列紧密,细胞基质分泌多,油红O染色后可见细胞内有大量脂滴形成。(B)茜素红染色后可见大量清晰的钙结节形成。结论:成功构建犬DFSCs膜片,并证明其具有较强的成骨能力,为进一步利用犬DFSCs膜片修复牙槽骨缺损的研究提供条件。Objective： To study the construction of dental follicle stem cell sheet and its biological characteristics in Beagle dogs.Methods： After identification,DFSCs were sub-cultured to construct DFSCs sheet.Cell sheet was investaged by inverted microscope,HE staining and scanning electron microscope（SEM）.DFSCs sheets were induced by adipogenesis inducing medium and osteogenic medium for 14 days separately.Oil red staining and Alizarin red staining was applied to examine adipogenic induction and osteogenic induction.Results： DFSCs showed typical spindle shape..Colony-forming assay results showed about 5.1% DFSCs colony formation.DFSCs were positive for CD29 and CD44,but negative for CD34.MTT manifested the growth and proliferation was good.Cell cycle testing showed： G1=87.1%,G2=5.54%.DFSCs sheets were constructed successfully and its growth in multilayer.It found that DFSCs expanded ade-quately and extracellular matrix（ECM） was clear and numerous in scanning electron micrescopy.Oil red staining and alizarin red staining both demonstrated positive reactions in DFSCs sheet after induction.Conclusion： It suggested that DFSCs cell sheet may be constructed and has a strong bone-forming ability.

关 键 词：牙囊干细胞(DFSCs) 细胞膜片 分化 

分 类 号：Q953[生物学—动物学] Q75