人子宫蜕膜树突状细胞DEC-205^+HLA-DR^+CD11c^+的分离纯化与鉴定

Separation and Culture of Decidual Dendritic Cells DEC-205^+HLA-DR^+CD11c^+ in Adult Uterus

目的:探讨人子宫蜕膜树突状细胞的分离方法和诱导培养的最佳条件,为研究树突状细胞在母胎免疫耐受中的作用奠定基础。方法:人早孕(7～12周)子宫蜕膜组织经机械破碎,梯度离心获得蜕膜低密度细胞,经粒-巨噬细胞集落刺激因子(GM-CSF)和干细胞因子(SCF)诱导,所获得细胞用FITC标记的抗人DEC-205、HLA-DR、CDllc抗体进行鉴定。结果:从蜕膜组织可获取足够数量蜕膜树突状细胞,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,20d内细胞数仍能维持在(1.0～2.3)×105个/mL。结论:梯度离心法能成功分离蜕膜树突状前体细胞,GM-CSF和SCF体外诱导可提供蜕膜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 decidual dendritic cells, and establish the ground work for the research of dendritic cell＇s functionary mechanism in maternal immunity and tolerance to the fetus. Methods The fresh early gestation （7-12 wk gestation） decidual tissue was dispersed mechanically in Hank＇S balanced salt solution, and the low density cells were obtained by gradient centrifugation and cultured with granulocytemacrophage colony-stimulating factor （GM-CSF） and stem cell factor （SCF）. Then the generated cells were identified by phenotypic and functional methods. Results A total of 1.0-2.3×10^5 decidual dendritic cells were obtained from the fresh decidual 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 decidual dendritic cells. GM-CSF and SCF can promote the growth of the decidual dendritic cells and provide satisfactory growth conditions for them.