脐血贴壁细胞体外培养与生长特性观察

The Growth Characteristics of Cultured Cord Blood Adherent Cells

目的观察脐血贴壁细胞生长特性及提高体外培养脐血贴壁细胞成功率,为造血干细胞培养提供基质环境。方法采用密度梯度离心法分离脐血单个核细胞,在含20ng/ml bFGF,10%胎牛血清的低糖DMEM培养基中培养,于倒置显微镜下观察细胞生长情况、并描绘其生长曲线、流式细胞仪检测细胞周期。结果脐血贴壁细胞呈长梭形、针形或三角形,增殖周期G0/G1为85.12%,倍增时间为48h。结论体外培养的脐血细胞能够贴壁生长并形成单层细胞层。

Objective To study the growth characteristics of cord blood adherent cells and provide the hematopoietic microenvironment. Methods Mononuclear cells were isolate from umbilical cord blood by density gradient centrifugation, then the cells were cultured in the L-DMEM medium with 20ng/ml bFGF, 10%FBS. The cells were observed by invert microscope. Drawing the growth curve and testing the cell cycles with flowing cytometry. Results Cord blood adherent cells display fibroblast like or spindle shape, and their reproductive cycle： G0/G1 was 85.12%, the doubling time was 48 hours. Conclusions The cord blood cells could adhere the flask and proliferate in vitro, at last form a single cell layer.