摘要
目的 分离培养并鉴定人胚胎骨髓间充质干细胞。方法 抽取流产胎儿股骨骨髓，梯度离心，取含有骨髓间充质干细胞的低密度血小板层进行培养；免疫细胞化学法鉴定细胞膜抗原和细胞基质中的蛋白属性；酶细胞化学法测定细胞碱性磷酸酶活性。结果 培养的细胞呈现纺锤形外观；细胞膜抗原 PDGFR、CD29、CD44有阳性表达，IGF-1R、CD34、CD45呈阴性表达，细胞基质中纤维粘连蛋白、胶原蛋白Ⅰ、胶原蛋白Ⅲ有表达，而胶原蛋白Ⅱ和层粘蛋白没有表达；碱性磷酸酶检测呈阴性反应。结论 培养的细胞不是骨髓中的造血干细胞或骨髓基质中的成纤维细胞，是处于未分化阶段的骨髓间充质干细胞。
Objective To culture and identify mesenchymal stem cells (MSCs) derived from human fetal bone marrow. Methods Human fetal bone marrow was obtained and gradient centrifugation performed to isolate cell pellets of low density that contained MSCs followed by cell culture. Immunocytochemistry technique was employed to examine the cell surface antigens and matrix enzymes, and enzyme cytochemistry used to detect alkaline phosphatase (ALPase). Results The cultured cells were characterized by spindle-shaped appearance and positive expression of PDGFR, CD29, CD44, fibronectin, collagen I and collagen Ⅲ were detected while IGF-1R, CD34, CD45, collagen Ⅱand laminin were negative. Enzyme cytochemistry evaluation showed that the cultured cells were negative for ALPase. Conclusion The cultured cells were not hemopoietic stem cells or fibroblasts but the MSCs that had not differentiated.
出处
《第一军医大学学报》
CSCD
北大核心
2001年第8期571-573,F002,共4页
Journal of First Military Medical University
基金
国家重点基础研究发展规划(973)(G1999054038)
国家自然科学基金项目(30070368)
广州市科委科技攻关项目(2000-Z-017- 01-05
