Lee JangBo, Kuroda Satoshi, Shichinohe Hideo, Ikeda Jun, Seki Toshitaka, Hida Kazutoshi, Tada Mitsuhiro, Sawada Ken-ichi, Iwasaki Yoshinobu
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Neuropathology. 2003 Sep;23(3):169-80. doi: 10.1046/j.1440-1789.2003.00496.x.
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 bis-benzimide 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.
越来越多的证据表明，骨髓基质细胞（BMSC）有潜力迁移至受损神经组织并分化为中枢神经系统（CNS）细胞，这表明自体移植治疗具有可能性。本研究旨在阐明，将小鼠BMSC移植到局灶性脑梗死或脊髓损伤的小鼠体内时，其是否能够迁移至损伤部位并分化为CNS细胞。从小鼠体内获取BMSC，并通过流式细胞术进行鉴定。然后，用核荧光染料双苯甲酰亚胺对BMSC进行24小时以上的标记，并通过立体定向技术将其移植到小鼠的脑或脊髓中。流式细胞术检测显示，培养的BMSC表达低水平的CD45和高水平的CD90及Sca-1。移植后4周，大量移植细胞在正常脑内存活，其中许多位于移植部位附近。荧光免疫组织化学检测显示，它们表达包括NeuN、微管相关蛋白2（MAP2）和双皮质素在内的神经元标志物。然而，当将BMSC移植到大脑中动脉闭塞小鼠的同侧纹状体时，许多移植细胞迁移至胼胝体和受损皮质，并且在移植后4周也表达神经元标志物。特别是，NeuN对于验证移植细胞的分化非常有用，因为该标志物在细胞核中表达且与双苯甲酰亚胺重叠。脊髓损伤的小鼠也得到了类似结果。然而，许多移植的BMSC在受损脊髓中表达星形胶质细胞蛋白胶质纤维酸性蛋白（GFAP）。本研究结果表明，小鼠BMSC能够迁移至CNS损伤部位并分化为神经元或星形胶质细胞，并且双苯甲酰亚胺是一种简单且有用的标志物，可用于标记供体细胞并长期评估其在宿主神经组织中的迁移和分化情况。
