单个核细胞和单核源性巨噬细胞生长特性的体外实验研究

Growth feature of mononuclear cell and monocyte-derived macrophage in vitro

目的研究人外周血单个核细胞、单核源性巨噬细胞在体外培养环境中的生长曲线情况。方法以密度梯度离心法和贴壁法分离、纯化正常人的外周血单个核细胞。以佛波醇酯促进单核源性巨噬细胞的分化成熟。用MTT法检测单个核细胞、单核源性巨噬细胞的生长曲线。采用单因素方差分析进行统计学检验。结果单个核细胞接种第1天的细胞数量明显多于第3、5、6、7天（P〈0.05）,但接种后第2~7天的细胞数量相对稳定（P〉0.05）。单核源性巨噬细胞分化成熟后第1~6天的OD,细胞数量较稳定。但第7天的细胞数量明显少于前6天（P〈0.05）。结论体外培养第2~7天的外周血单个核细胞处于生长平台期。体外培养的单核源性巨噬细胞在分化成熟后的第1~6天处于生长平台期。

Objective To explore the growth curve of mononuclear cells or monocyte-derived macrophage in vitro.Methods Ficoll gradient centrifugation was used to obtain mononuclear cells and remove non-adherent lymphocytes.Mononuclear cells were cultured with PMA（100nmol/L） to obtain monocyte-derived macrophage.Cellular growth curve was measured by MTT tests.Results The quantity of mononuclear cells was stable from 2th to 7th day after inoculation.The quantity of monocyte-derived macrophage was stable from 1th to 6th day after maturity.Conclusion The plateau of mononuclear cells and monocyte-derived macrophage is 2-7th days in vitro and 1-6th days after mature,respectively.