人重症肌无力胸腺树突状细胞的分离和培养  

Separation and culture of thymic dendritic cells in adult myasthenia gravis thymus

作　　者：刘苹[1] 赵云平[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所分子生物学中心,重庆400042 [2]第三军医大学大坪医院野战外科研究所全军胸外科中心,重庆400042

出　　处：《中华实验外科杂志》2004年第1期31-32,共2页Chinese Journal of Experimental Surgery

摘　　要：目的　探讨人重症肌无力胸腺树突状细胞 (TDC)的分离方法和诱导培养的最佳条件 ,为研究胸腺树突状细胞的生理功能及其在重症肌无力抗原提呈中的作用奠定基础。方法　新鲜人胸腺组织经机械破碎 ,梯度离心获得胸腺低密度细胞 ,经粒 巨噬细胞集落刺激因子 (GM CSF)和干细胞因子 (SCF)诱导 ,所获得细胞通过透射电镜、扫描电镜观察 ,并用FITC标记的抗人S10 0b、HLA DR、CD11c抗体进行鉴定。结果　从胸腺组织可获取足够数量胸腺树突状细胞 ,体外培养 10～ 18d细胞生长较为活跃 ,第 14天左右达到最高值 ,维持 3～ 4d开始下降 ,2 0d内细胞数仍能维持在 (1.0～ 2 .5 )× 10 5个 /ml。结论　梯度离心法能成功分离胸腺树突状前体细胞 ,GM CSF和SCF体外诱导可提供胸腺树突状细胞原代培养较为满意的生长条件。Objective To investigate the separation methods and growth conditions of thymic dendritic cells (TDC),and establish the ground work for the research of TDC physiologic function and antigen-presenting in myasthenia gravis.Methods The fresh thymic tissue was dispersed mechanically in Hank's balanced salt solution,and the low density cells were obtained by gradient centrifugation and cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and stem cells factor (SCF).Then the generated cells were identified by ultrastructural,phenotypic and functional methods.Results A total of 1.0～2.5×10 5 thymic dendritic cells were obtained from the fresh thymic tissue.These cells grew vigorously within 10～18 days and reached the peak at the 14th day.Conclusion The method of mechanically,gradient centrifugation can successfully separate the TDC.GM-CSF and SCF can promote the growth of the TDC and provide satisfactory growth conditions for them.

关 键 词：重症肌无力 胸腺树突状细胞 分离 培养 

分 类 号：R746.1[医药卫生—神经病学与精神病学]