胎源性非黏附骨髓基质细胞的特性

Characteristics of fetal non-adherent bone marrow stromal cells

背景:骨髓中存在非黏附干细胞,该类细胞形态及表型特征与黏附骨髓间充质干细胞相似。目的:观察胎源性非黏附骨髓基质细胞的基本特性。方法:采用反复贴壁分离法培养胎源性骨髓非黏附基质细胞,每24h取悬浮细胞分瓶培养,共4次;采用MTT法测定第1,3,5代细胞生长曲线,流式细胞仪行细胞周期测定和表面标志分析;并取第1代细胞行造血集落培养,第3代细胞成骨和成脂诱导培养。结果与结论:反复贴壁分离培养4次后仍有细胞贴壁并有集落形成,剩余细胞经培养7d后即可得到原代骨髓非黏附基质细胞。骨髓非黏附基质细胞形态不均匀,高表达整合素及内皮细胞表面标记,中等量表达CD106,同时也表达造血细胞标记CD34;体外可分化为成骨细胞及造血细胞,但成脂分化能力较弱。

BACKGROUND： There are non-adhesion stem cells in bone marrow stem cells, and the morphology and phenotype characteristics of the cells are similar to adhesion bone marrow mesenchymal stem cells. OBJECTIVE： To explore the characteristics of fetal non-adherent bone marrow stromal cells （NA-BMSCs）. METHODS： The fetal NA-BMSCs were cultivated by the method of adherent separation repeatly （＂pull-offs＂）, the suspended cells were incubated in another culture bottle every 24 hours for 4 times. The growth curve of the passage 1, 3, 5 NA-BMSCs was detected by MTT assay, and the phenotype of the NA-BMSCs was determined with flow cytometry. Passage 1 NA-BMSCs were preformed with hematopoietic colony assay and passage 3 NA-BMSCs were preformed with osteogenic and adipogenic induction. RESULTS AND CONCLUSION： After four times’ ＂pull-off＂, there were still asherent cells and colony formation; the resulting cells were continuing to culture for 7 days and those cells were the primary NA-MSCs （P0）. The morphology of the cells was uneven; they highly expressed the integrin and the surface markers of endothelial cells with a middle level of CD106. Interestingly, they also expressed the surface markers of hematopoietic cells CD34. Those cells had multi-differentiation potency in vitro and they could differentiate into osteoblasts and hematopoietic cells under different inducing medium, but the capability of lipid differentiation was weak.