免疫磁珠分选的基质细胞抗原1(STRO-1)阳性人牙周膜细胞具有多向分化的干细胞特性  

Multiple differentiation potential of human periodontal ligament stem cells expressing stromal cell antigen 1(STRO-1)sorted by MACS

作　　者：赵萤 屠腾 丰帆 张旻 ZHAO Ying;TU Teng;FENG Fan;ZHANG Min(State Key Laboratory of Military Stomatology,National Clinical Research Center for Oral Diseases,Shaanxi International Joint Research Center for Oral Diseases,Department of General Dentistry&Emergency,School of Stomatology,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病国际联合研究中心,空军军医大学口腔医学院急诊与综合临床科,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2020年第11期971-976,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(31971248,81901052)。

摘　　要：目的采用免疫磁珠法(MACS)分选人牙周膜细胞基质细胞抗原1阳性(STRO-1^+)细胞及STRO-1^-细胞,并对两种细胞的干细胞特性进行对比分析。方法组织消化法分离培养人牙周膜细胞,并采用STRO-1抗体结合的免疫磁珠对细胞进行分选。随后使用流式细胞仪检测STRO-1^+细胞和STRO-1^-细胞中表面标志物CD90、CD105、CD29、STRO-1、CD34和CD45的表达;采用集落形成实验检测分离细胞的集落形成率;在成骨和成脂诱导分化后,分别采用茜素红染色和油红O染色鉴定其多向分化能力。结果STRO-1^+细胞可高表达干细胞表面标志物CD90和CD105,而低表达造血细胞标志物CD34和CD45;并且STRO-1^+细胞的集落形成率显著高于STRO-1^-细胞;STRO-1^+细胞具有向成骨和成脂细胞分化的潜能。结论采用免疫磁珠法分选的STRO-1^+细胞为具有干细胞特性的人牙周膜干细胞。Objective To perform in vitro characterization and comparison of stromal cell antigen 1(STRO-1)positive(STRO-1^+)and negative(STRO-1^-)cell populations from human periodontal ligament cells sorted by magnetic-activated cell sorting(MACS).Methods Periodontal ligament cells were isolated from the periodontal ligament by tissue digesting method.The cells were magnetically sorted using anti-STRO-1 antibodies.Subsequently,cells were characterized by flow cytometry for the expression of CD90,CD105,CD29,STRO-1,CD34 and CD45.Colony-forming assay was used to detect the ability of colony-forming.When the cells are induced to differentiate by osteogenesis and lipogenesis,alizarin red staining,and oil red O staining were used to identify the pluripotent differentiation of STRO-1^+and STRO-1^-cells.ResultsSTRO-1^+cells highly expressed CD105 and CD90 and lowly expressed the haematopoietic markers CD45 and CD34.In contrast to STRO-1^-cells,STRO-1^+cells showed a significantly higher number of colony-forming units(CFUs).STRO-1^+cells were demonstrated to have osteogenic and adipogenic differentiation potential.Conclusion STRO-1^+cells isolated by MACS are human periodontal ligament stem cells with stem cell characteristics.

关 键 词：牙周膜干细胞 牙周膜 细胞分选 免疫磁珠法 

分 类 号：R392.11[医药卫生—免疫学] R392-33[医药卫生—基础医学]