Migration and differentiation of nuclear fluorescence-labeled bone marrow stromal cells after transplantation into cerebral infarct and spinal cord injury in mice
Migration and differentiation of nuclear fluorescence-labeled bone marrow stromal cells after transplantation into cerebral infarct and spinal cord injury in mice
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DOI：
10.1046/j.1440-1789.2003.00496.x
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发表时间：
2003-09-01
期刊：
影响因子：
2.3
通讯作者：
Iwasaki, Y
中科院分区：
文献类型：
--
作者：
Lee, JB;Kuroda, S;Iwasaki, Y
越来越多的证据表明，骨髓基质细胞(BMSC)具有向受损神经组织迁移和分化为中枢神经系统细胞的潜能，提示自体移植治疗的可能性。本研究旨在阐明小鼠骨髓间充质干细胞移植到局灶性脑梗塞或脊髓损伤小鼠体内后，是否能迁移到病变部位并分化为中枢神经系统细胞。取小鼠骨髓间充质干细胞，用流式细胞仪对其进行鉴定。然后，用双苯甲亚胺标记骨髓间充质干细胞，并将其立体定向移植到小鼠的脑或脊髓中。流式细胞仪检测显示，BMSC低水平表达CD45，高水平表达CD90和SCA-1。移植后4周，正常脑组织内有大量移植细胞存活，其中许多细胞位于移植部位附近。荧光免疫组织化学法检测神经元标志物NeuN、MAP2、Doublecortin的表达。而将骨髓间充质干细胞移植到大脑中动脉闭塞小鼠同侧纹状体内，移植后4周，移植的细胞大量迁移至胼胝体和损伤皮质，并表达神经元标志物。特别是，NeuN对于验证移植细胞的分化非常有用，因为该标记表达在细胞核中，并与双苯甲亚胺重叠。在脊髓损伤的小鼠身上也得到了类似的结果。然而，许多移植的BMSC在损伤的脊髓中表达星形细胞蛋白GFAP。结果表明，小鼠骨髓间充质干细胞可向中枢神经系统损伤处迁移并向神经元或星形胶质细胞分化，双苯甲亚胺是一种简便、实用的标记供体细胞的方法，可用于长期观察供体细胞在宿主神经组织中的迁移分化情况。
There is increasing evidence that bone marrow stromal cells (BMSC) have the potential to migrate into the injured neural tissue and to differentiate into the CNS cells, indicating the possibility of autograft transplantation therapy. The present study was aimed to clarify whether the mouse BMSC can migrate into the lesion and differentiate into the CNS cells when transplanted into the mice subjected to focal cerebral infarct or spinal cord injury. The BMSC were harvested from mice and characterized by flow cytometry. Then, the BMSC were labeled by bis-benzimide, a nuclear fluorescence dye, over 24 h, and were stereotactically transplanted into the brain or spinal cord of the mice. The cultured BMSC expressed low levels of CD45 and high levels of CD90 and Sca-1 on flow cytometry. A large number of grafted cells survived in the normal brain 4 weeks after transplantation, many of which were located close to the transplanted sites. They expressed the neuronal marker including NeuN, MAP2, and doublecortin on fluorescent immunohistochemistry. However, when the BMSC were transplanted into the ipsilateral striatum of the mice subjected to middle cerebral artery occlusion, many of the grafted cells migrated into the corpus callosum and injured cortex, and also expressed the neuronal markers 4 weeks after transplantation. In particular, NeuN was very useful to validate the differentiation of the grafted cells, because the marker was expressed in the nuclei and was overlapped with bis-benzimide. Similar results were obtained in the mice subjected to spinal cord injury. However, many of the transplanted BMSC expressed GFAP, an astrocytic protein, in injured spinal cord. The present results indicate that the mouse BMSC can migrate into the CNS lesion and differentiate into the neurons or astrocytes, and that bisbenzimide is a simple and useful marker to label the donor cells and to evaluate their migration and differentiation in the host neural tissues over a long period.