一种高效的巨噬细胞体外诱导培养方法

A novel high-yield macrophage culture in vitro

[目的]探索一种不需要分离前体细胞和加入外源性细胞因子的巨噬细胞体外诱导培养方法。[方法]收集腹腔接种肝癌腹水瘤细胞的小鼠腹水细胞,37℃、5%CO2条件下不换液、不传代体外培养。当培养细胞几乎全部死亡时,有少量细胞的胞浆内颗粒消失、折光性变强。此时少量换液,这些细胞的体积迅速增大、分裂增殖活跃。通过形态学观察、免疫学标志检查及体外吞噬功能检测,鉴定这些细胞是巨噬细胞。[结果]本实验获得了高纯度、高活性的巨噬细胞,具备巨噬细胞的形态特征和巨噬细胞特异的分化抗原-CD68。与不同诱导方法培养的巨噬细胞的吞噬能力比较,差异均有显著性意义(P<0.05),表明死亡细胞诱导的巨噬细胞具有良好的吞噬能力。[结论]应用不需要分离前体细胞和加入外源性细胞因子的巨噬细胞体外诱导培养方法,操作简便、经济实用,易于获得足量的巨噬细胞,可广泛应用于临床及实验研究。

[ Objective I To explore a method of macrophage induced cultivation in vitro without precursor cell and exogenous cytokine. [ Methods] Mouse ascites cells were collected from peritoneal cavity and cultivated without replacing culture medium and passing era in vitro at the condition of 37℃ and 5% CO2. When the whole cultivated ceils nearly died, a few ceils showed that granules in plasma disappeared and refraction became strong. These cells were rapidly augmented and turned in- to proliferation immediately after replacing little culture medium. By morphological obsepcation, immunological symbol examination and phagocytic function detection in vitro ,these cells were identified as macrophages. [ Results ] Macrophages with high activity were purified according to morphologic characteristics and immunological symbol of specific differentiation antigen of macrophage - CD68. Macrophages induced by death cells had notable discrepancy compared to macrophages that were cultivated by other methods （P 〈 0.05 ）. It showed that macrophages induced by death cells had well phagocytic abili- ty. [ Conclusion I This is the first time to use method of macrophage induced cultivation in vitro without precursor cell and exogenous cytokine. It is a simple, economic and practical method for obtaining sample macrophages applied in clinical and experimental research broadly.