微囊化基质细胞对脐带血造血干/祖细胞扩增支持

将脐带血单个核细胞与包埋有兔骨髓间充质干细胞的海藻酸钙微胶珠在3种不同的培养液中进行了7d的体外静态共培养.每24h进行总有核细胞计数,在0、72和168h进行流式CD34+细胞分析以及甲基纤维素集落检验.实验结果表明:经过7d的静态共培养,在添加常规剂量造血生长因子的培养液中,总有核细胞扩增了(15±2.85)倍,CD34+细胞扩增了(5.33±0.32)倍,CFU-Cs扩增了(5.6±1.21)倍.微胶囊可以作为一种新的共培养隔离手段,微囊化兔骨髓间充质干细胞在添加适量血清或者造血生长因子组合的条件下对于脐带血造血干/祖细胞在静态下的扩增有明显的促进作用.

Cord blood hematopoietic stem/progenitor cells expansion supported by encapsulated stromal cells

The coculture of cord blood mononuclear cells (CB MNCs) and alginate beads containing rabbit bone marrow (BM) mesenchymal stem cells (MSCs) was investigated in three kinds of culture mediums under the static condition for 7 days. The total nucleated cell density was measured every 24 hours. Flow cytometric assay for CD34~+ cells and methylcellulose colony assays were carried out at 0,72 and 168 h. The results show that the total nucleated cell number is multipled by (15±2.85) times,CD34~+ cells (5.33±0.32) times and colony-forming units in culture (CFU-Cs) (5.6±1.21) times respectively at the end of the 7th day's coculture in the serum-free medium supplemented with conventional dose of cytokines. Microcapsules may be the appropriate coculture protocol and the encapsulated rabbit BM MSCs illustrate remarkable effects on CB hematopoietic stem/progenitor cells expansion when serum or proper cytokines are added under the static condition.