Objective: To analyze the proliferation potential of bone marrow-derived mesenchymal stem cells (MSC) in patients with myelodysplastic syndrome (MDS).
Methods: The MSC derived from the 24 patients with newly diagnosed MDS (MDS-MSC group) and MSC derived from 15 patients with nutritional anemia (control group) in the Affiliated Hospital of Hebei University were used as the research objects. The proliferation potential of MSC was analyzed by colony-forming unit assay, doubling time, cumulative passaging, cell number after 10 days of culture with equal amount of MSC and MTT experiment. The mechanism of abnormal proliferation was analyzed by cell cycle experiment, apoptosis experiment and p21 gene expression assay.
Results: In the colony forming unit assay, the number of MDS-MSC colonies was 4.44±2.51, which was significantly lower than that of the control group (12.44±2.55)(P<0.01); the doubling time of MDS-MSC group was significantly longer than that of the control group (7.80±3.26 vs 3.63±0.85) (P<0.01); the number of MDS-MSC in 5×104 culture for 10 days was (39.40±14.18)×104, which was significantly lower than that of the control group ［(85.30±9.49)×104 ］(P<0.01); the number of cumulative passages in MDS-MSC group was 5.20±1.40, which was significantly lower than that in control group (11.60±1.96)(P<0.01); MTT results showed that the proliferation capability of MSC in MDS-MSC group was lower than that in the control group. The cell proportion of G0/G1 phase in MDS-MSC group was higher than that in the control group, while the cell proportion of S phase was lower (P<0.05). The percentage of early apoptotic cells in MDS-MSC group was higher than that in control group (P<0.05); the relative expression level of p21 mRNA in MDS-MSC group was significantly higher than that in control group(P<0.01).
Conclusion: The proliferative capability of MDS-MSC is significantly reduced, which relates with the arrest of cell cycle in G0/G1 phase, the increase of early apoptotic cells and senescent cells of the MDS-MSC.
题目: 骨髓增生异常综合征患者来源的骨髓间充质干细胞增殖特点的分析.
目的: 分析骨髓增生异常综合征（MDS）患者骨髓来源间充质干细胞（MSC）的增殖特点.
方法: 以在河北大学附属医院就诊的24例初治MDS患者的MSC（MDS-MSC 组）和15例营养性贫血患者的MSC（对照组）为研究对象，通过CFU实验、计算MSC的倍增时间、累积传代数、等量MSC培养10 d后的细胞数和MTT实验分析MSC的增殖能力；通过细胞周期实验、细胞凋亡实验及p21基因的表达分析MDS-MSC增殖能力改变的潜在原因.
结果: 在CUF实验中，MDS-MSC的集落数(4.44±2.51)明显低于对照组(12.44±2.55)（P＜0.01）； MDS-MSC的倍增时间为(7.80±3.26) d，显著长于对照组MSC［(3.63±0.85) d］（P＜0.01）； 5×104培养10 d后MDS-MSC数量为(39.40±14.18)×104，低于对照组 ［(85.30±9.49) ×104］（P＜0.01）； MDS-MSC的累积传代数为5.20±1.40，显著低于对照组（11.60±1.96）（P＜0.01）；MTT实验结果显示，MDS-MSC增殖活力低于对照组MSC。MDS-MSC组中G0/G1期的细胞比例高于对照组，同时S期的细胞比例下降（P＜0.05）；MDS-MSC的早期凋亡细胞数高于对照组MSC（P＜0.05）；MDS-MSC组中p21的mRNA表达水平显著高于对照组MSC（P<0.01）.
结论: MDS-MSC增殖能力显著下降，这与MDS-MSC阻滞在G0/G1期、早期凋亡细胞数和衰老的细胞比例升高有关.