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目的研究从G-CSF动员的外周血细胞(PBC)悬液培养的成纤维细胞样(F-L)细胞的特性。方法取PBC中贴壁细胞分4组培养①RPMI-1640组;②L-DMEM组;③粒细胞集落刺激因子(G-CSF)组;④白细胞介素-3(IL-3)组。流式细胞仪分析各组培养的F-L细胞特性。结果培养2-3周后,4组均能收获到F-L细胞,细胞贴壁生长,但不融合,不能连续传代培养,③④组细胞数量明显多于①②组,4组F-L细胞表型相似CD33+、CD11c+、CD64+、CD14+、CD45+、HLA-DR+、CD86+、CD34-、CD38-、CD3-、CD19-、CD56-、CD29-、CD44-、CD105-;与单核细胞(PB-M)表型差异仅CD38表达不同而与间质干细胞(MSC)或树突状细胞(DC)表型明显不同。结论从PBC培养的F-L细胞为巨噬细胞,不是MSC或DC;G-CSF、IL-3能提高F-L细胞培养数量,不改变PB-M向巨噬细胞分化的分化方向。
Objective To investigate the characteristics of fibroblast-like (F-L) cells cultured from G-CSF-mobilized peripheral blood (PBC) suspensions. Methods Adherent cells in PBC were divided into 4 groups: ①RPM-1640 group; ②L-DMEM group; ③G-CSF group; ④interleukin-3 (IL-3) group. The characteristics of F-L cells cultured in each group were analyzed by flow cytometry. RESULTS: After 2-3 weeks of culture, FL cells were harvested in all four groups. However, they did not fuse, and could not be subcultured continuously. The number of cells in ③④ group was significantly more than that in ①② group. The FL cell phenotypes of the four groups were similar to that of CD33 + CD11c +, CD64 +, CD14 +, CD45 +, HLA-DR +, CD86 +, CD34-, CD38-, CD3-, CD19-, CD56-, CD29-, CD44-, CD105-; Differences in expression of CD38 alone are significantly different from that of mesenchymal stem cells (MSCs) or dendritic cells (DCs). CONCLUSION: F-L cells cultured from PBC are macrophages, not MSC or DC. G-CSF and IL-3 can increase the number of F-L cells cultured without changing the differentiation direction of PB-M into macrophages.