外周血MSCs的分离培养及对G-CSF的动员反应

The feasibility analysis of MSCs mobilization by G-CSF and its prosthetic effection in traumatic brain injury

目的建立多种种属外周血间充质干细胞(MSCs)的分离培养方法,探讨其对粒细胞集落刺激因子(G-CSF)的动员反应。方法用percoll(1.131g/mL)分离多种种属外周血单个核细胞,进行贴壁培养MSCs。以G-CSF作为动员剂,培养骨髓和外周血来源的MSCs,计数动员前后的成纤维细胞集落形成单位(CFU-F)。结果对多种种属进行外周血MSCs分离培养,成功率不同。在本实验条件下,可以稳定的直接分离培养出大鼠外周血的MSCs。G-CSF动员后,骨髓来源MSCs的CFU-F数量显著高于对照组(P<0.01);外周血来源MSCs的CFU-F数量也显著高于对照组(P<0.01),接近4倍。结论不同种属外周血MSCs分离培养难易不同,其直接原因是生理条件下外周血中存在的MSCs量少导致的。G-CSF可以有效地动员大鼠骨髓和外周血MSCs。

Objective To establish the isolating cultural method of MSCs from some genus peripheral blood （PB） , and explore the mobilization response of MSCs by G-CSF. Methods MSCs from some genus PB were isolated by percoll （ 1. 131 g/mL） and carried out adherent culture. MSCs-derived bone marrow and PB were cultured and its CFU-F were counted after mobilization by G-CSF. Results PB MSCs were isolated and cultureed successfully or unsuccessfully from different genus. Under our experiment condition, rat＇ s PB MSCs could been isolated and cultured steadily. A significantly higher number of CFU-F of BM-derived MSCs and PB-derived MSCs than that of control group after mobilization by G-CSF（ P 〈 0.01 ） ,ahuost 4-fold increase. Conclusion PB MSCs were isolated and cultured successfully or unsuccessfully from different genus, it concerned with a small number of circulating MSCs in BP. We confirmed that G-CSF mobilized BM-derived MSCs, especially PB-derived MSCs by CFU-F assay.