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

Time Effect of Proliferating Cell Nuclear Antigen Antisense Oligodeoxynucleotide on the Expansion and Differentiation of Umbilical Cord Blood CD34^+ Hematopoietic Precursor Cells in vitro

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72 h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。

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^＋ CD386- cells were expanded （55.1 ± 4.2） folds, and （49.8 ± 2.5） % cells were in G0/G2 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.