体外菲立磁标记恒河猴骨髓基质细胞自体移植入纹状体后磁共振对标记细胞形态学的示踪观察

Tracking observation of cell morphology with magnetic resonance after autologous transplantation of bone marrow stromal cells of rhesus monkey into the striatum labeled with Feridex in vitro

目的:体外菲立磁标记的骨髓基质细胞经单细胞悬液微移植后,观察将其在恒河猴纹状体的存活、迁移、分化和整合状况,为细胞移植治疗疾病奠定基础。方法:实验于2004-04/2005-05在南方医科大学珠江医院全军神经医学研究所和解放军第四军医大学神经科学研究所完成。纳入健康恒河猴3只,离恒河猴骨髓基质细胞,利用神经干细胞培养基、白血病抑止分因子和碱性成纤维母细胞生长因子进行细胞扩增并诱导成骨髓基质源神经干细胞,再经体外菲立磁和活细胞荧光染料PK67标记后,采用微移植的方法,通过脑立体定位仪上用微玻璃针将干细胞分别植入脑纹状体内。细胞移植后1,4,8周应用核磁共振成像对脑内移植的细胞进行活体示踪,最后利用光镜和电镜观察标记细胞在脑内的形态学情况。结果:①体外菲立磁标记结果:骨髓基质细胞经微移植后可在脑内纹状体区域存活,移植的干细胞可向周围的脑实质内迁移和整合,迁移细胞沿特定的纹状体结构分布,少量细胞可分化成神经元。②核磁共振成像检查结果:发现脑内移植的标记细胞在磁共振上呈明显的低信号改变,未标记细胞侧脑组织无明显的低信号改变,与组织学切片结果基本相一致。结论:髓基质源神经干细胞移植后,可在脑内存活、迁移、分化和整合,骨利用核磁共振成像技术可以对脑内移植后的标记细胞进行初步的活体追踪。

AIM： To investigate the survival, migration, differentiation and incorporation of labeled bone marrow stromal cells （BMSCs） of rhesus monkey after being transplanted into the striatum with single cell suspension and to investigate the possibility of a new cell source for central nervous system transplantation. METHODS： The experiment was conducted at the Institute of Military Neurology Medical, Zhujiang Hospital, Southern Medic＇,：d University and Institute of Neuroscience, Fourth Military Medical University of Chinese PLA from April 2004 to May 2005. Three heahhy rhesus monkeys were involved. BMSCs of rhesus monkey were separated and performing cell amplification and induce into stem cells of BMSCs-D-NSCs with NSCs euhure medium, leukemia inhibitor and basic fibroblast growth factor. After these cells were further labeled with Feridex and PKH67 in vitro, they were transplanted into striatum by microtransplantation method with brain solid positioner. After 1, 4 and 8 weeks, the monkeys were traced in vivo and their brain tissues were sectioned to investigate the morphology of the transplanted cells under light microscopy and electron microscopy.RESULTS： ①The results of Feridex showed that BMSCs could survive in striatum of brain after microtransplantation, Stem cells transplanted could migrate and incorporate into the host brain tissue and were distributed along special striatum structure, but a few of them differentiated into neurons. ②MRI showed remarkably low signal change was seen in the left brain transplanted with labeled BMSCs and no signal change in contralateral brain. MR images were basically correlated with histopathological section result. CONCLUSION： The transplanted BMSCs can survive, migrate, differentiate and incorporate in the brain. MRI technique is ideally suited for primary in vivo tracking of labeled stem cells after grafting.