摘要
目的从人脐血中分离、培养间充质干细胞(MSCs)并探讨其对淋巴细胞的免疫调节作用。方法淋巴细胞分离液分离人脐血单个核细胞,利用贴壁筛选法通过多次传代得到MSCs,流式细胞仪测定细胞表型;将获得的MSCs分别以不同数量加入到外周血混合淋巴细胞培养体系和植物血凝素(PHA)刺激的外周血淋巴细胞转化体系中,用H3-TdR标记β液体闪烁计数仪检测细胞增殖,观察脐血来源的MSCs对混合淋巴细胞反应和淋巴细胞转化的影响。结果从人脐血中分离获得的贴壁细胞,呈成纤维样的细胞形态,CD29、CD105和CD166表达阳性,CD14、CD34和CD45表达阴性;人脐血源性MSCs在体外对混合淋巴细胞反应和PHA诱导的淋巴细胞转化均具有明显的抑制作用,抑制作用与细胞数量呈正相关。结论从人脐血中可以成功地分离出MSCs,其对同种异体淋巴细胞具有免疫调节作用,为其作为骨组织工程异基因种子细胞来源打下基础。
Objective To study the isolation, expansion, and purification of mesenchymal stem cells （MSCs） from human umbilical cord blood, and investigate its immune regulatory effects on homologous lymphocyte. Methods Human umbilical cord blood samples were obtained sterilely. The cord blood mononuclear cells were isolated by lymphocyte separation medium and individual colonies were selected and cultured. The phenotype of MSCs derived from umbilical blood was analyzed by flow cytometry. The MSCs derived from umbilical blood were added to mixed lymphocyte cultures and PHA-induced transformation cultures with various concentrations. The proliferation of lymphocyte was measured by [^3H]-thymidine incorporation using a liquid scintillation counter. Effects of MSCs derived from umbilical blood on mixed lymphocyte response and PHA-induced lymphocyte transformation were investigated. Results The adherent cells isolated from the cord blood mononuclear cells displayed a fibroblast-like morphology and expressed several MSCs-related antigens CD29, CD44, and CD105, not CD34, CD45, or CD106. The MSCs derived from umbilical blood showed negative immunomodulatory effects and could inhibit evidently the mixed lymphocyte response and PHA induced transformation. Conclusion The MSCs derived from umbilical blood with the immune regulatory effect could be an important and novel source of allogenic seed cells for bone tissue engineering.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第3期287-290,294,共5页
Immunological Journal
基金
国家高技术研究发展计划重大攻关课题资助("863"计划2006AA02A122)
关键词
脐血
间充质干细胞
免疫调控
骨组织工程
Umbilical cord blood
Mesenchymal stem cells
Immunomodulation
Bone tissue engineering
