K562细胞红系诱导分化方法的实验研究

Experimental Research on the K562 Cells Erythroid Differentiation

目的建立一种高效K562细胞红系诱导分化方法。方法比较不同方法对K562细胞红系诱导分化的效果,优化诱导剂成分组合和剂量组合,建立一种高效的红系诱导,鉴定指标选用细胞形态学、联苯胺染色阳性率计数及RT-PCR测定红系细胞标志物膜带3蛋白(band3)的表达。结果多因素协同诱导K562细胞红系分化120 h后,显微镜下观察细胞形态呈红系中、晚期特征,联苯胺染色阳性率可达(52.7±8.2)%,红系标志物band3在mRNA水平明显高于对照组(P<0.01)。结论多条信号途径协同参与K562细胞红系分化的过程,多因素共同诱导可显著提高K562细胞的红系分化。

Objectlve To establish an efficient induction of K562 cells erythroid differentiation. Methods The effect of different inducing methods of K562 cells erythroid differentiation were compared, the compositions and dose were optimized to establish a highly efficient erythroid induction method, the identification indicators were cell morphology and benzidine staining positive rate count, further exploring of erythroid differentiatiott was performed by RT-PCR to test the cell membrane marker band3 protein （band3） expression. Results The percentage of benzidine-positive ceils induced by the optimized condition was （52.7 ± 8.2）% at 120 hours; RT-PCR showed the mRNA level of band3 was significantly higher than the non-induced group（ P 〈 0.01 ）. Conclusion Several synergistic signaling pathways are involved in erythroid differentiation of K562 ceils, multi-factors co-induction can significantly enhance K562 cells erythroid differentiation.