人骨髓间充质干细胞表面抗原单克隆抗体的制备及初步鉴定

Preparation Monclonal Antibody of Human Mesenchymal Stem Cell

目的制备人骨髓间充质干细胞(MSCs)的特异性细胞表面抗原的单克隆抗体。方法以人骨髓MSCs为抗原,常规方法免疫Balb/c小鼠,细胞融合制备杂交瘤细胞,并用免疫荧光法在流式细胞仪上对杂交瘤产生的抗体进行初测和复测。结果通过分泌的抗体与人MSCs表面抗原特异性结合的鉴定,进行杂交瘤筛选,将复测为阳性的克隆连续克隆化培养,获得两株持续分泌抗人MSCs的杂交瘤细胞株:898、8f9。两株单抗经体外连续传代培养,生长良好,稳定分泌抗体。结论人骨髓MSCs的特异性细胞表面抗原单克隆抗体可望在MSCs的提纯和体外富集、特异性骨病的诊断等方面有广阔的应用前景。

Objective To find monclonal antibody on cell surface of mesenchymal stem cell (MSCs) from human bone marrow. Methods Immuned Balb/c rice with MSCs were derived from human bone marrow, then hybridizated the immuned spleen with SP 2/0 cells. For evaluation of the hybridoma cells, immumofluorescence method with flow cytometer were used. Results Hybridoma cells were with standard that whether they can secret antibody which has specificty conjunction with MSCs sufarce antigen. After successive culture of the hybridoma cells which are positive in re-evaluta-tion, two stain hybridoma cells was found:8g8, 8f9. The antibody secreted from the two belongs to IgG subclass. Conclusion It is expected that the monoclonal antibody will be applied in many fields, such as treatment and diagnosis.