摘要
目的阐明小鼠骨髓基质细胞体外培养的影响因素,优化骨髓基质细胞体外培养方法。方法以小鼠骨髓基质细胞为研究对象,倒置显微镜和MTT法观察骨髓基质细胞不同接种数目(1×105个/孔,3×105个/孔,5×105个/孔)条件下和相同接种数目不同培养基体积(100、150、200μl)条件下的骨髓基质细胞体外生长状态。制备骨髓基质细胞体外培养条件培养基,观察条件培养基对骨髓基质细胞体外生长状态的影响。结果骨髓基质细胞体外培养7 d,100μl培养体系时,接种1×105个细胞的培养孔基本无基质细胞生长,接种3×105个细胞的基质细胞贴壁约30%左右,接种5×105个细胞的基质细胞贴壁达90%左右。当接种细胞数目为3×105个时,随培养基体积的增多基质细胞数目明显减少(P<0.05),而条件培养基组基质细胞的数目明显多于普通培养基组(P<0.05);当接种细胞数目为5×105个时,培养基体积对基质细胞生长状态基本无影响,条件培养基组与普通培养基组骨髓基质细胞的生长状态无显著差异。结论骨髓基质细胞在体外培养时,接种细胞数目过低会影响自身分泌胞外因子的浓度从而影响其自身的生长;细胞自分泌对自身的生长有促进作用,但是当分泌的胞外因子达到一定浓度后促生长作用将达到平台期。
【Objective】To elucidate the factors influencing the growth of murine bone marrow stromal cells in vitro and to optimize the culture method of these cells.【Methods】Murine bone marrow cells were used as cell model.The growth state of bone marrow stromal cells was observed in vitro by inverted microscope and MTT assay when the cells were inoculated with different cell number（1 × 105/well, 3 × 105/well, 5 × 105/well） and with the same cell number but different media volume（100, 150, 200 μl）. The conditioned medium was prepared, and the effect of conditioned medium was observed on growth of bone marrow stromal cells in vitro. 【Results】After bone marrow cells were cultured for 7 days in vitro, the adherent cells grew well and reached about 90% as 5 × 105 cells /well were inoculated, the adherent cells reached about 30% as 3 × 105 cells /well were inoculated, but the adherent cells could not be found as 1 × 105 cells /well were inoculated. When the number of inoculated cells was 3 × 105cells/well, with the increase in the volume of the medium, the number of adherent cells was significantly reduced（P 0.05）, and the conditioned medium significantly promoted the growth of bone marrow stromal cells（P 0.05）. When the number of inoculated cells was 5 × 105 cells /well, the volume of the medium and the conditioned medium had no effect on the growth of bone marrow stromal cells.【Conclusions】Bone marrow stromal cells can secrete extracellular factors to promote their own growth in vitro, and the proliferative effect can reach a plateau when the extracellular factors reach a certain concentration.
出处
《中国现代医学杂志》
CAS
北大核心
2015年第21期17-20,共4页
China Journal of Modern Medicine
基金
国家"重大新药创制"科技重大专(No:2009ZX09103-708)
国家自然科学基金(No:81403317)
广东省自然科学基金(No:1015102240100002)
广东省学科建设专项资金项目(No:2013LYM 0044)
关键词
骨髓基质细胞
自分泌
细胞生长
bone marrow stromal cell
autocrine
cell growth
