M21170应用粒细胞集落刺激因子体外扩增脂肪基质干细胞

Application of granulocyte colony-stimulating factor to proliferate adipose stromal cells in vitro

目的探讨粒细胞集落刺激因子(granulocyte colony stimulating factor,G-CSF)对大鼠脂肪基质干细胞(adipose stromal cells,ASCs)体外扩增能力的影响。方法取10只SD大鼠腹膜后脂肪组织,差异贴壁法分离培养ASCs。检测第3代ASCs常规培养与加入G-CSF条件下体外扩增能力与细胞周期分布情况。结果①实验组ASCs形态与对照组比较无明显变化。②实验组大鼠ASCs培养18d后,细胞集落数目(14.9±4.6),细胞总数(3.9±0.8)×104;对照组细胞集落数目(7.7±3.8),细胞总数(2.4±0.6)×104。实验组与对照组比较,ASCs细胞集落数目与细胞总数差异均有统计学意义(P<0.05)。③实验组ASCs细胞G0-1细胞比例相对常规培养组低(P<0.05)。结论G-CSF可促进ASCs进入细胞增殖周期,增加其体外扩增能力。

[Objective] To apply granulocyte colony-stimulating factor to proliferate Adipose stromal cells （ASCs） in vitro. [Methods] The ASCs of ten rats were separated and cultured by the discrepancy-adherence method. The ASCs （P3） of every rats were cultivated in vitro by two different ways： conventional culture and culture with G-CSF （100μg/L） for 18 days. For ASCs of all rats： The shape of ASCs received different culture mothed were observed. The clone number and the total cellular score of ASCs were observed in vitro. The generation cycle of ASCs were investigated with the flow cytometry. [Results] The shape of ASCs received conventional culture have no difference from that of the ASCs received culture with G-CSF. The clone number, which were （14.9±4.6）, and the total cellular score of ASCs received culture with G-CSF, which were （3.9±0.8）×10^4, were significant different from that of ASCs received conventional culture, which respectively were （7.7±3.8） and （2.4±0.6）×10^4. The cell cycle distribution of ASCs received conventional culture had significant difference with ASCs received culture with G-CSF. [Conclusion] G-CSF can stimulate ASCs rentering cell cycle and promote the proliferation of ASCs.