经侧脑室注射人脐带间充质干细胞向胶质瘤趋化能力的实验研究

The experimental investigation of glioma-trophic capacity of human umbilical cord-derived mesenchymal stem cells after intraventricular administration

目的探讨经侧脑室注射人脐带间充质干细胞向胶质瘤的趋化迁移能力。方法无菌条件下采集足月妊娠剖宫产患者的正常脐带组织(知情同意),经胰酶和胶原酶消化、贴壁培养获得人脐带间充质干细胞,通过细胞形态学观察、流式细胞术分析,以及向脂肪细胞、成骨细胞和神经细胞的多向分化潜能证实其间充质干细胞之特征。将CM-DiI标记的人脐带间充质干细胞注射至C6胶质瘤荷瘤大鼠病灶对侧侧脑室,2周后观察其趋瘤能力和在瘤床、肿瘤与正常脑组织交界处、"卫星"瘤灶内分布情况。结果人脐带间充质干细胞呈均匀一致的纤维母细胞样贴壁生长,分别表达CD13、CD29、CD44和CD90等间充质干细胞标志物,而不表达CD14、CD31、CD34、CD38、CD45、CD133等造血和内皮细胞标志物以及CD49、CD106和HLA-DR。经诱导分化培养后,人脐带间充质干细胞可分化为脂肪细胞、成骨细胞、神经元样和星形胶质细胞样细胞,表明其具有多向分化潜能。CM-DiI标记的人脐带间充质干细胞经侧脑室注射后可向胶质瘤定向迁移,分布于瘤床、肿瘤与正常脑组织交界处和"追踪"浸润至正常脑实质的"卫星"瘤灶。结论人脐带间充质干细胞具有向胶质瘤特异性靶向迁移并在瘤内广泛分布的能力,可作为胶质瘤基因治疗的理想细胞载体。

Objective To explore the glioma-trophic migration capacity of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs) by intraventricular administration.Methods The umbilical cord tissue were obtained during full-term pregnancy cesarean section under sterile conditions.This study was approved by Ethics Committee and got the informed consent of patient.The hUC-MSCs were isolated by trypsin and collagenase digestion,followed by adherent culture methods.The characteristics of isolated hUC-MSCs were demonstrated by cell morphylogy,phenotype analysis and multi-differentiation potentials into adipocytes,osteoblasts and neural cells.Then the hUC-MSCs were labeled with CM-DiI and injected into contralateral ventricle of glioma of the C6 glioma-bearing Sprague-Dawley(SD) rats.Two weeks later,the rats were sacrificed and the brains were taken out to examine the migration and distribution of hUC-MSCs in the tumor bed,at the interface of tumor and cerebral parenchyma as well as the tumor satelites infiltrating into the normal brain.Results The hUC-MSCs demonstrated plastic-adherent characterization and homogeneous fibroblastic-like morphylogy in culture,expression of specific surface phenotypes of MSCs(CD13,CD29,CD44,CD90) but not endothelial or hematopoietic markers(CD14,CD31,CD34,CD38,CD45,CD133),and muti-differentiatiation potentials into Oil red O stained adipocytes,Alizarin red S stained osteoblasts,neuron-specific enolase(NSE)-positive neurons and glial fibrillary acidic protein(GFAP)-positive astrocytes in permissive inducive conditions.Importantly,after labeled hUC-MSCs injection into contralateral ventricle of glioma,the hUC-MSCs migrated from initial injection site to the glioma mass and along the interface of tumor and brain,and some of them 'chasing' the glioma satellites infiltrated into the normal parenchyma.Conclusion The hUC-MSCs possess prominent tumor-specific targeting capacity and extensive intratumoral distribution in glioma models.Thus,they may serve as novel vehicles in cell-based gene-therapy of glioma.