论文部分内容阅读
目的研究人骨髓基质细胞体外长期培养的生物学特性和造血支持功能。方法①采用静置贴壁细胞培养法，体外长期培养胎儿、儿童和成人的骨髓基质细胞。②采用免疫细胞化学染色法和流式细胞仪检测法，分析细胞的表型。③将不同发育阶段的骨髓基质细胞体外培养，并扩增脐血造血干细胞。结果①建立了成纤维肌样细胞系，可传至１０代，维持６个月，同时还培养出内皮细胞和巨噬细胞。②儿童骨髓基质肌样细胞的染色特征为：波形纤维蛋白（ｖｉｅｍｅｎｔｉｎ）呈阳性，第Ⅷ因子呈阴性；儿童骨髓基质细胞的表型为：ＣＤ３３－、ＣＤ３４－、ＣＤ３８－和ＣＤＷ９０＋；成人为：ＣＤ３３－、ＣＤ３４－、ＣＤ３８＋和ＣＤＷ９０＋。③基质细胞能长期维持脐血中ＬＴＣ－ＩＣ的生存。若加入ＩＬ－６、ＩＬ－３和ＳＣＦ扩增体系，基质细胞对长期维持和扩增ＬＴＣ－ＩＣ的效果更为明显（Ｐ〈０．０１），较无基质细胞的对照组所形成的ＬＴＣ－ＩＣ产率高２－４倍。结论人骨髓基质细胞体外可长期培养，对脐血干细胞具有造血支持功能。
Objective To study the biological characteristics and hematopoietic function of human bone marrow stromal cells cultured in vitro for a long time. Methods ① Adopting static adherent cell culture method to culture fetal, children and adult bone marrow stromal cells in vitro for a long time. ② using immunocytochemical staining and flow cytometry, analysis of cell phenotype. ③ different stages of bone marrow stromal cells in vitro culture, and expansion of cord blood hematopoietic stem cells. Results ① The fibroblast-like cell line was established, which could be passaged to passage 10 for 6 months and endothelial cells and macrophages were also cultured. ② The staining characteristics of bone marrow stromal myoblasts in children were: positive for viementin and negative for factor Ⅷ; the phenotypes of children with bone marrow stromal cells were: CD33-, CD34-, CD38- and CDW90 +; adults were: CD33-, CD34-, CD38 +, and CDW90 +. ③ stromal cells can long-term maintenance of cord blood LTC-IC survival. When added to IL-6, IL-3 and SCF, the effect of stromal cells on long-term maintenance and expansion of LTC-IC was more obvious (P <0.01) than that of control cells without stromal cells -IC high yield 2-4 times. Conclusion Human bone marrow stromal cells can be cultured in vitro for long time and have hematopoietic function of cord blood stem cells.