增殖细胞核抗原反义寡核苷酸对人脐血CD34+细胞分化的影响

目的:研究增殖细胞核抗原反义寡核苷酸（PCNA-ASODN）对脐血造血干/祖细胞体外扩增的调节作用。 方法:采用免疫磁珠分离技术从新鲜脐带血中分离CD34+细胞，使用不同浓度的PCNA-ASODN作用于体外扩增的CD34+细胞，应用流式细胞技术检测各种细胞数量，细胞内PCNA表达及细胞周期。 结果:加入了PCNA-ASODN的两个实验组，其PCNA的表达率为（27.2±3.6）%和(19.0±1.5)%，低于对照组的（53.8±8.3）%（P<0.01）。低浓度组扩增的细胞中CD34+细胞比例为（33.4±3.2）％，显著高于对照组的（25.2±2.6）%（P<0.01），而且CD34+CD38-细胞数量达到（57.8±9.9）倍，也显著高于对照组的（43.5±7.4）倍(P<0.01)。 结论:低浓度的PCNA-ASODN有效抑制CD34+细胞在增殖过程中PCNA表达，减缓扩增过程中的分化作用，提高了早期祖细胞的扩增效率。

Effect of proliferating cell nuclear antigen specific antisense oligonucleotide on differentiation of cord blood CD34+ cells

AIM:To study the effect of proliferating cell nucler antigen antisense oligonucleotide on ex vivo expansion of cord blood CD34+ hematopoietic stem/progenitor cells. METHODS:CD34+ cells were purified from fresh cord blood by immunomagnetic beads. CD34+ cells were incubated in liquid culture system with different concentrations of PCNA-ASODN. Using flow cytometry, the number of different kinds of stem/progenitor cells and PCNA expression were measured after CD34+ cell incubation. RESULTS:PCNA was lowly expressed in low experiential group, with a positive rate of (27.2±3.6)% and (19.0±1.5)%, the positive rate of control group was (53.8±8.3)% (P<0.01), a high significant difference was observed. Just in low concentration of PCNA-ASODN, the percentage of CD34+cells were increased to (33.4±3.2)%, CD34+CD38-cells expanded (57.8±9.9) folds, and the percentage of CD34+cells were (25.2±2.6)% (P<0.01), the CD34+CD38-cells expanded (43.5±7.4) folds (P<0.05), it has significant difference compared with the control group. CONCLUSION:Low concentration of PCNA-ASODN decreases PCNA expression effectively, slows down differentiation of CD34+cells during ex vivo expansion procedure, and improves the expansion efficiency.