小鼠脾脏成纤维网状细胞的分离、培养及鉴定

Isolation,cultivation and identification of fibroblastic reticular cells from the spleen stromal cells

目的建立小鼠脾脏基质细胞培养的方法 ,分离出成纤维网状细胞并对其进行鉴定。方法取健康C57小鼠脾脏,胶原酶消化后,用磁珠分选出CD45-脾脏基质细胞,接种于α-MEM培养基,传代培养待其稳定生长。流式细胞仪进一步分选出成纤维网状细胞(gp38+CD31-),并用PCR、免疫荧光的方法检测其趋化因子表达。结果利用磁珠分离出脾脏的基质细胞培养后可以稳定传代,成功的分选出小鼠脾脏成纤维网状细胞(gp38+CD31-),并检测到其CCL19和CCL21的表达。结论成功分离培养脾脏基质细胞,并分选出有功能的成纤维网状细胞,为实验研究提供稳定的细胞来源。

Fibroblastic reticular cells（FRCs） form a network in the T cell zone（periarteriolar lymphoid sheath,PALS） of the spleen white pulp（WP） on which are important for homing T lymphocytes and dendritic cells. In this study,spleen stromal cells were enriched by CD45 magnetic beads separation,and were cultured in α-MEM high glucose complete medium. Flow cytometry was applied to identify and sort the FRCs. Immunofluorescence staining and PCR were conducted to identify expression of CCL21 and CCL19. Sorting by flow cytometry,we isolated the FRCs（gp38^＋CD31^-） from the spleen stromal cells successfully,and these cell express CCL21 and CCL19.