间充质干细胞协同BMP-2蛋白对造血干细胞增殖的影响

The effect of bone marrow mesenchymal stem cell synergistic BMP-2 in the proliferation of hematopoietic stem cell

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3 d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。

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 m RNA and protein expression of Ki67 were detected by fluorescence q RT-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.