阵发性睡眠性血红蛋白尿症患者骨髓CD_(34)^+CD_(59)^+细胞的体外扩增

Ex vivo expansion of CD_(34)^+CD_(59)^+ cells from bone marrow of paroxysmal nocturnal hemoglobinuria patients

目的 研究阵发性睡眠性血红蛋白尿症 (PNH)患者CD3 4 + CD59+ 细胞的分离、纯化及其体外扩增的条件和性能 ,为探索PNH新的治疗途径提供实验依据。方法 利用免疫磁珠 流式细胞仪二步分选法 ,从PNH患者骨髓中分选出CD3 4 + CD59+ 细胞 ,然后对CD3 4 + CD59+ 细胞在不同造血生长因子组合条件下 ,进行体外扩增培养 2周。结果 体外扩增的最适宜的生长因子组合为SCF +IL 3+IL 6+FL +Tpo +Epo,最适宜的扩增时机为第 7天 ,在此条件下 ,CD3 4 + CD59+ 细胞的扩增倍数为 2 2 .4 2±3 73倍。CD3 4 + CD59+ 细胞在扩增以后 ,仍保持较好的形成集落形成单位的能力 ,但是其向多系分化的潜能有所下降。结论 PNH患者CD3 4 + CD59+ 细胞能够进行体外扩增。按照最佳扩增条件 ,在对PNH患者进行自体骨髓移植或自体外周血干细胞移植时有一定的应用价值。

Objective To study the separation, purification and ex vivo expansion of CD 34 +CD 59 + cells from patients with paroxysmal nocturnal hemoglobinuria(PNH), and explore the new treatment for the PNH patients. Methods CD 34 +CD 59 + cells were selected from the bone marrow mononuclear cells of PNH patients by means of immunomagnetic microbead flow cytometry two step sorting method, followed by ex vivo expansion of the cells with combination of hematopoietic factors for two weeks. Results The best combination for the ex vivo expansion was SCF+IL 3+IL 6+FL+Tpo+Epo, and the seventh day was the most suitable time for the best harvest when the CD 34 +CD 59 + cells were 22.42±3.73 fold expanded. After ex vivo expansion, the cells remained CD 59 positive and potent capacity of colony formation, but their potentialities to multilineage differentiation were decreased. Conclusion The present study shows that ex vivo expansion of CD 34 +CD 59 + cells from PNH patients might promise the possibility of performing ABMT or APBSCT clinincally for the patients.