培养基对兔骨髓间充质干细胞体外扩增的影响

Proliferation in vitro of bone marrow-derived mesenchymal stem cells in different culture meeia in rabbits

目的考察含10%胎牛血清的高糖DMEM培养基(DMEM-HG,10%FBS)、10%胎牛血清的低糖DMEM培养基(DMEM-LG,10%FBS)、15%胎牛血清的低糖DMEM培养基(DMEM-LG,15%FBS)、20%胎牛血清的低糖DMEM培养基(DMEM-LG,20%FBS)对兔骨髓间充质干细胞体外贴壁、扩增的影响。方法选2月龄的新西兰大白兔,股骨转子间区抽取骨髓利用密度梯度离心贴壁法分离纯化细胞,体外培养。选取传代第一代骨髓间充质干细胞(passenger one,P1)分别用以上4种细胞培养基培养,测细胞扩增倍数;②选取传代第二代骨髓间充质干细胞(pas-senger two,P2)分别将以上4种细胞培养基混悬液接种于24孔板中,观察细胞生长状态并测生长曲线;③选取4种培养基中培养效果最好的一组细胞,以密度0.5×104/mL接种于24孔板中,成骨诱导,茜素红染色测其矿化能力。结果DMEM-LG(15%FBS)组中细胞扩增倍数为(16.20±1.60)倍,细胞集落形成数为6.11±1.17,与其它组比较差异有统计学意义(P<0.05),并矿化能力最强。结论DMEM-LG(15%FBS)培养基较其他3种培养基更符合兔骨髓间充质干细胞体外扩增培养的条件,且更好的保持了细胞的正常形态和生物活性。

Objective To investigate the effects of different culture media on the proliferation in vitro of bone marrow-derived mesenchymal stem cells. Methods Bone marrow was abstracted from the intertrochanter of the femur of 2-month New Zealand rabbits and purification of mesenchymal stem cells was determined by a density gradient centrifugation method. The cells were cultured in 4 kinds of ctdture media： DMEM-high glucose（ 10% FBS）, DMEM-low glucose（ 10% FBS）, DMEM-low glucose （15%FBS） and DMEM-low glucose（20%FBS）. Cells of passage one were detected the expansion multiple and cells of passage two measured the growth and mineralization of cells. Results The expansion multiple was 16.20 ±1.60 and the colony-forming number（CFN） was 6.11± 1.17 in the DMEM-low glucose （ 15 % FBS） group, which were significantly higher than those in the other three groups. Conclusion DMEM-low glucose （15% FBS） is more suitable for cultivation of MSCs than the other three media.