大鼠骨髓间充质干细胞培养、鉴定、SPIO标记及MRI活体示踪的实验研究

发布时间：2018-01-31 04:39

本文关键词： 脑创伤骨髓间充质干细胞超顺磁性氧化铁颗粒示踪磁共振成像　出处：《福建医科大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的分离、培养、鉴定及SPIO标记大鼠BMSCs,探讨MRI对其在大鼠脑创伤模型活体示踪成像中的可行性。方法 1.全骨髓贴壁培养法分离培养大鼠BMSCs,流式细胞仪检测细胞表面抗原及向成脂、成脂诱导分化进行鉴定,并探讨首次换液模式对BMSCs培养时间及细胞纯度的影响。 2.以左旋多聚赖氨酸(PLL)为转染介质介导导BMSCs吞噬SPIO,评价SPIO对细胞生物学特性的影响,探讨最佳SPIO浓度。 3.参照Feeney法制作大鼠TBI模型,经右侧经总动脉穿刺插管注射移植SPIO标记的BMSCs。 4.实验组大鼠注射移植SPIO标记BMSCs,对照组TBI大鼠注射等量的无SPIO标记的BMSCs或PBS,均分别于注射细胞前及注射BMSCs后1d、3d、7d、14d、21d进行NSS评分及MRI扫描,观察脑实质内信号变化及其迁移情况,并与对侧正常脑实质信号进行对比。每次MRI扫描结束后立即各处死1只,取脑组织切片进行HE及普鲁士蓝染色。结果 1.BMSCs生长状态良好,培养时间较短,纯度较高,CD29、CD90阳性,CD34、CD45阴性,向成骨、成脂诱导分化,SOIO标记后细胞生物活性良好。 2.MRI扫描可观察到实验组TBI模型大鼠脑实质内低信号区,并逐步向病变区迁移,与对侧脑实质内信号减低率结果具有统计学意义(F=154.19,P0.05),病理切片普鲁士蓝染色见蓝染颗粒。BMSCs移植组NSS评分低于PBS移植组,结果有统计学意义(P0.05)。结论 1.全骨髓贴壁法培养BMSCs生物学特性良好,可被SPIO有效标记且不影响生物活性。 2. 3.0T MRI联合动物专用线圈对SPIO标记的BMSCs活体示踪有意义。 3.BMSCs可能对大鼠神经功能恢复有一定作用。
[Abstract]:Purpose To investigate the feasibility of MRI in the in vivo tracer imaging of rat brain trauma model, we isolated, cultured, identified and labeled BMSCs with SPIO. Method 1. Whole bone marrow adherent culture method was used to isolate and culture BMSCs. Flow cytometry was used to detect cell surface antigen and adipogenic differentiation. The effects of the first liquid exchange model on the culture time and cell purity of BMSCs were investigated. 2. The phagocytosis of BMSCs mediated by L-polylysine (PLL) was studied to evaluate the effect of SPIO on cell biological characteristics and to explore the optimal concentration of SPIO. 3. The rat model of TBI was made by referring to the method of Feeney, and the BMSCs labeled by SPIO were injected into the right common artery via the right side. 4. The experimental group was injected with SPIO labeled BMSCs, while the control group with TBI was injected with the same amount of BMSCs or PBS without SPIO labeling. NSS scores and MRI scans were performed before the injection of the cells and 1 day after BMSCs injection. The changes of signal and migration in the brain parenchyma were observed. At the end of each MRI scan, 1 rat was killed and the brain sections were stained with HE and Prussian blue. Results 1. BMSCs grow well, culture time is short, the purity of BMSCs is high, CD29 + CD90 + CD34 + CD34 CD45 negative, osteogenesis, adipogenic differentiation. SOIO labeled cells showed good bioactivity. 2. The hypointense area in the brain parenchyma of the experimental group TBI model rats was observed by MRI scanning, and the brain moved to the lesion area gradually. The results of signal reduction in the contralateral cerebral parenchyma were statistically significant (P 0.05). Prussian blue staining of pathological sections showed that the NSS score of blue dye granules. BMSCs transplantation group was lower than that of PBS transplantation group, and the result was statistically significant (P 0.05). Conclusion 1. The biological characteristics of BMSCs cultured by whole bone marrow adherent method were good, which could be effectively labeled by SPIO and had no effect on biological activity. 2.3.0T MRI combined with animal coil has significance for SPIO labeled BMSCs tracer in vivo. 3. BMSCs may play a role in the recovery of neural function in rats.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332

【参考文献】