摘要
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。
Aim To study on biological properties and hemopoiesissupporting function of human bone marrow stromal cells cultured for long-term in vitro. Methods ① Bone marrow stromal cells from fetus, chidren and adults were cultured for long-time in vitro by static adherent wall assay. ② The phenotypes of the stromal cells were analyzed by immunocytochemical staining or FACA. ③ Hemopoietic stem cells in umbilical cord- blood were co-cultured and expanded with the stromal cells at different development stages. Results ① The fibroblast myoid cell lines were established and could be propagated to 10 generations and kept for 6 months, meantime, endothelial cells and macrophoges were obtained. ② The child stromal myoid cells were positive for viementin and negative for Ⅷ factor by immunocytochemical staining and their phenotypes were CD33 -, CD34 -, CD38 - and CDW90+, while the phenotypes of adult stromal cells were CD33-, CD34-, CD38+ and CDW90+. ③ The stromal cellls could support long-time survival or expansion of LTC-ICs. Survival the of LTC-ICs in the stromal cell-supporting system supplemented with SCF, IL-3 and IL-6 was even longer than that in system without stromal cells(P<0. 01 ). The productivity rate of LTC-ICs cultured in the stromal cell-supporting system was 2 to 4 times higher than that cultured in system without stromal cells. Conclusion The human bone marrow stromal cells could be cultured for long-time in vitro and have hemopoiesis-supp ortingfunction for the hemopoietic stem cells in cord blood.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第1期41-45,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
江苏省自然科学基金资助!No.BJ99507
关键词
骨髓基质细胞
造血支持功能
生物学特性
细胞培养
bone marrow
stromal cells
cord blood
long-term culture-initiating cell
hemopoiesis-supporting founction
