摘要
采用静置贴壁细胞培养法，体外长期培养了胎儿、儿童、成人骨髓基质细胞，时间为６个月，可传至１０代，并观察了成纤维肌样细胞、内皮细胞和巨噬细胞；通过免疫细胞化学染色法，证明了１～３代的骨髓基质肌样细胞Ｖｉｅｍｅｎｔｉｎ为阳性，第Ⅷ因子阴性；采用流式细胞仪检测法，证明了儿童骨髓基质细胞的细胞表型为ＣＤ３３－和ＣＤ３４－、ＣＤ３８－、ＣＤ＋Ｗ９０，成人骨髓基质细胞为ＣＤ－３３、ＣＤ－３４、ＣＤ＋３８、ＣＤ＋Ｗ９０。 采用体外培养和扩增的脐血造血干细胞的半固体集落形成法，证明了基质细胞能长期维持脐血中的ＬＴＣ－ＩＣ的生存，若加入ＩＬ－６、ＩＬ－３、ＳＣＦ细胞因子的扩增体系，基质细胞对长期维持和扩增ＬＴＣ－ＩＣ的效果更为明显（Ｐ＜０．０１），比无基质细胞的对照组所形成的ＬＴＣ－ＩＣ产率高于２～４倍。表明骨髓基质细胞具有支持造血功能。
Fetal, child, adult bone marrow stomal cells were cultured for six months in vitro by static adherent assay, and propagated up .to 10 times, The myoid fibroblast, endothelial and macrophagic cells were observed. The result of immunocyte chemisty intricated the myoid cells propagated 1 to 3 times were positive for viementin and negtive forⅧ factor. The result of phenotype analysis by FACS suggested the child bone marrow stromal cells were CD33-、CD34-、 CD38-、 CDW90+, and adult cells were CD33、CD34、CD38、CDW90 The CFC assay was used to test the hematopoietic in cold bleed after culture and expansion in vitro, the result demonstrated the long - term supportive effect of stromal cells on the survival of LTCICs in cord blood. The expansion of LTC-ICs in the stromal-supported system containing SCF, IL-3,IL-6 was more remarkable than that in nonstroma system (P< 0.01). The frequency of LTC - ICs of the stromal - supported system was 2 to 4 times higher than that of nonstroma system. The results above suggested that the bone stromal cells have bone marrow hematopoiesis - supported function.
出处
《中国血液流变学杂志》
CAS
2000年第2期87-93,共7页
Chinese Journal of Hemorheology
基金
江苏省自然科学基金
关键词
人骨髓基质细胞
造血支持作用
细胞培养
Human bone marrow stromal cells Long- term culture- initiating cells (LTC - ICs) Fibroblast Hematopoiesis - supported function
