增殖细胞核抗原反义寡脱氧核苷酸调节脐血CD34+造血前体细胞体外扩增和分化的时间效应_临床医学论文.docVIP

增殖细胞核抗原反义寡脱氧核苷酸调节脐血CD34+造血前体细胞体外扩增和分化的时间效应_临床医学论文 增殖细胞核抗原反义寡脱氧核苷酸调节脐血CD34+造血前体细胞体外扩增和分化的时间效应_临床医学论文 【摘要】 目的:探讨低浓度增殖细胞核抗原反义寡脱氧核苷酸(PCNA_ASODN)调节脐血造血前体细胞体外扩增和分化的时间效应。方法:用免疫磁珠分离技术从新鲜脐带血中分离出CD34+细胞,先后加入低浓度PCNA_ASODN作用于体外培养的CD34+细胞,用流式细胞仪检测培养后各种细胞的数量及细胞周期的变化。结果:实验组脐血CD34+造血前体细胞明显被扩增，但以第72h组的扩增效应最为显著，其CD34+细胞比例增高［(35.6±1.8)%］,而CD34+CD38－细胞数量扩增达(55.1±4.2)倍，且G0/G1期细胞占(49.8±2.5)%。结论:低浓度PCNA_ASODN对脐血CD34+造血前体细胞体外扩增的调节具有时间效应,同时能延缓扩增过程中伴随的细胞分化。 【关键词】 增殖细胞核抗原反义寡脱氧核苷酸 造血前体细胞 体外扩增 　　［Abstract］Objective: To study the time effect of low concentration proliferating cell nuclear antigen antisense oligodeoxynucleotide(PCNA_ASODN)on the expansion and differentiation of umbilical cord blood CD34+　hematopoietic precursor cells in vitro. Methods: CD34+　cells were purified from fresh umbilical cord blood by immunomagnetic beads, and incubated in liquid culture system with low concentration PCNA_ASODN which added into the culture system at different time. Using flow cytometry, the number of different kinds of stem/progenitor cells and cell cycle were detected after CD34+　cells incubated for 7 days. Results: The umbilical cord blood CD34+ hematopoietic precursor cells in the experimental group were obviously expanded, especially in the 72 hours group. The percentage of CD34+　cells in the 72 hours group was increased to(35.6±1.8)%, CD34+CD38－　cells were expanded(55.1±4.2)folds, and(49.8±2.5)% cells were in G0/G1 phase. Conclusion: Low concentration PCNA_ASODN has the time effect on the expansion of CD34+　cells, and reduces its differentiation when CD34+　cells are expanded in vitro. 　　［Key Words］proliferating cell nuclear antigen antisense oligodeoxynucleotide; hematopoietic precursor cell; expansion in vitro 　　近年脐血干细胞移植在治疗血液肿瘤和骨髓衰竭综合征方面取得了巨大成就，但由于单份脐血所能获得的造血干/祖细胞数量有限,限制了其在临床的广泛应用。脐血前体细胞的体外扩增是解决这一难题的有效途径之一，但扩增的同时不可避免地伴随细胞的分化成熟,从而影响了其重建长期造血的能力。如何在扩增造血前体细胞的同时尽可能地保留其自我更新能力和多向分化潜能，成为当前另一个研究热点。研究表明,造血前体细胞的增殖分化过程不但受控于细胞因子,同时与细胞周期调控因子密切相关［1,2］。增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)是重要的细胞周期调控