促血小板生成素对小鼠骨髓间充质干细胞动员的影响

Effect of Thrombopoietin on the Mobilization of Bone Marrow Mesenchymal Stem Cells

目的探讨不同时间取材,促血小板生成素(thrombopoietin,TPO)对小鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)动员的影响。方法将30只昆明小鼠(35±5)g随机分为TPO组与对照组,每组15只。TPO组腹腔注射TPO50μg/kg,对照组腹腔注射生理盐水0.1 ml/g,均连用5d。每组分别于最后一次注射后6 h、12 h、168 h收集并计数骨髓有核细胞(bone marrow nucleated cell,BMNC)和外周血有核细胞(peripheral blood nucleated cell,PBNC),同时以10~6/cm^2 BMNC、10~5/cm^2PBNC的密度分别接种,培养并计数成纤维细胞集落形成单位(colony forming unit-fibroblast,CFU-F)。结果与对照组相比,TPO组BMNC、PBNC细胞数分别增加(21.25~63.50)×10~5/ml,(4.40~12.46)×10~5/ml(P<0.05);TPO组骨髓来源CFU-F集落数增加了3~9倍(P<0.05),以12h所获集落数最多;在所有的外周血培养中,只有6 h和12 h TPO组观察到CFU-F样集落形成,但两组之间无统计学差异。结论 TPO动员小鼠骨髓MSCs的能力有限,与取材时间点无必然关系。

Objective To investigate the effect of thrombopoietin on the mobilization of bone marrow mesenchymal stem cells（MSCs）in mice at different time-points.Methods A total of 30 Kunming mice with mean weight（35±5）g were randomly divided into experimental and control groups（n=15in each group）.The experimental group was subjected to intraperitoneal injections of thrombopoietin（TPO）at a dose of 50μg/kg and the control group was treated with saline at a dose of 0.1 ml/g per day for 5 days.The bone marrow nucleated cell（BMNC）and peripheral blood nucleated cell（PBNC）were harvested on 6 h,12 h and 168 h after the final administration.The BMNC and PBNC were seeded at a density of 10~6cells/cm^2 and 10~5cells/cm^2 respectively.The colony forming unit-fibroblast（CFU-F）of MSCs was cultured and evaluated.Results Compared with control group,the number of BMNC and PBNC increased（21.25-63.50）×10~5/ml and（4.40-12.46）×10~5/ml in TPO groups and the number of BM-derived CFU-F increased 3-9 fold（P0.05）and harvested most on 12 h.The PB-derived CFU-F was only observed at 6h and 12 h in TPO group,but there was no significant difference between two groups.Conclusion There is no inevitable relationship between the limited ability of TPO on mobilizing MSCs in bone marrow to peripheral blood and Sampling time.