间充质干细胞协同BMP-2蛋白对造血干细胞增殖的影响

摘要

Objective To explore the proliferation effect of bone morphogenetic protein (BMP)-2 on hematopoietic stem cell (HSC) cocultured with mesenchymal stem cell (MSC).Methods MSCs were expanded to the third passage (P3) in the plastic dish;CD34+ cells were sorted by MACS Miltenyi Biotec.MSCs(P3) and HSCs were identified by FCM separately,and then MSC (P3) and HSC were co-cultured in the Transwell interfered with BMP-2 protein,four groups in the study:HSC group,HSC+BMP2 group,HSC+MSC group,HSC+MSC+BMP2 group.Cells were collected after 3 d to test HSC proliferation using the following methods:counting of the number,detecting of the total RNA,the mRNA and protein expression of Ki67 were detected by fluorescence qRT-PCR and immunofluorescence methods.Results The number of HSC,totel RNA,Ki67 expression of the HSC+BMP2 group was higher than HSC group,HSC+MSC group was higher than HSC group,HSC+MSC+BMP2 group was higher than HSC+MSC group (P ＜ 0.05).Conclusion MSCs synergistic with BMP-2 can enhance MSC proliferation effect on HSC.%目的 研究BMP-2联合人骨髓间充质干细胞(MSC)造血干细胞(HSC)体外增殖的影响.方法 培养MSC至第三代,磁珠分选仪分选HSC,并用流式细胞术鉴定MSC与HSC.将获得的MSC与HSC利用Transwell非接触共培养,100 ng/mL的骨形态发生蛋白(BMP-2)干预,即实验分四组:HSC组、HSC+BMP2组、HSC+MSC组、HSC+MSC+BMP2组.培养3d后比较不同培养条件下HSC计数、RNA浓度的不同,并通过实时荧光定量PCR法及免疫荧光方法检测HSC的Ki67的mRNA及蛋白的表达.结果 HSC的计数、总RNA含量、Ki67的表达在HSC+BMP2组高于HSC组、HSC+MSC组高于HSC组、HSC+MSC+BMP2组高于HSC+MSC组(P＜0.05).结论 MSC协同BMP-2蛋白可促进HSC的增殖.