移植到大鼠脊髓气压伤模型中的骨髓基质细胞的体内荧光追踪
In vivo fluorescence tracking of bone marrow stromal cells transplanted into a pneumatic injury model of rat spinal cord.
作者信息
Yano Shunsuke, Kuroda Satoshi, Lee Jang-Bo, Shichinohe Hideo, Seki Toshitaka, Ikeda Jun, Nishimura Goro, Hida Kazutoshi, Tamura Mamoru, Iwasaki Yoshinobu
机构信息
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
出版信息
J Neurotrauma. 2005 Aug;22(8):907-18. doi: 10.1089/neu.2005.22.907.
Recent experimental studies have shown that bone marrow stromal cells (BMSC) differentiate into neural cells and reduce neurological deficits when transplanted into traumatized spinal cord. These findings have been derived primarily from histological analyses. We conducted a study directed chiefly at developing a non-invasive system for tracking BMSC transplanted into the spinal cord of living animals. In this study, we induced spinal cord injury (SCI) in rats with a pneumatic device. BMSC were harvested from transgenic mice expressing green fluorescence protein (BMSC-GFP), and were transplanted stereotactically into a control group of rats without SCI (n = 6) and a group with SCI (n = 3). At 2 and 4 weeks after transplantation, the dura mater was exposed and green fluorescence derived from the transplanted BMSC-GFP was observed. The distribution and differentiation of the transplanted cells were subsequently evaluated with immunohistochemistry. Green fluorescence could be detected around the transplantation site in three of six of the control rats. In all three rats subjected to SCI, green fluorescence was shown to spread from the site of BMSC-GFP injection toward the injury site, suggesting that the transplanted cells had migrated toward the lesion within the 4-week post-transplantation period. Histological evaluation suggested that the detected green fluorescence was emitted by cells that had distributed in the dorsal white matter, and demonstrated that some of the transplanted cells expressed neuronal or astrocytic markers. These results suggest the possibility of tracking BMSC transplanted into the spinal cord in living animals. Such noninvasive bioimaging techniques would be valuable for monitoring the fate of these transplanted cells and assessing the safety and efficacy of their transplantation.
最近的实验研究表明,骨髓基质细胞(BMSC)移植到受创伤的脊髓后可分化为神经细胞并减少神经功能缺损。这些发现主要来自组织学分析。我们开展了一项主要旨在开发一种非侵入性系统的研究,用于追踪移植到活体动物脊髓中的BMSC。在本研究中,我们用气动装置诱导大鼠脊髓损伤(SCI)。从表达绿色荧光蛋白的转基因小鼠中获取BMSC(BMSC-GFP),并通过立体定位将其移植到未发生SCI的对照组大鼠(n = 6)和发生SCI的大鼠组(n = 3)中。移植后2周和4周,暴露硬脑膜,观察到移植的BMSC-GFP发出的绿色荧光。随后用免疫组织化学评估移植细胞的分布和分化情况。在6只对照大鼠中的3只,移植部位周围可检测到绿色荧光。在所有3只发生SCI的大鼠中,绿色荧光显示从BMSC-GFP注射部位向损伤部位扩散,这表明移植细胞在移植后4周内已向损伤部位迁移。组织学评估表明,检测到的绿色荧光是由分布在背侧白质中的细胞发出的,并证明一些移植细胞表达神经元或星形胶质细胞标志物。这些结果提示了在活体动物中追踪移植到脊髓中的BMSC的可能性。这种非侵入性生物成像技术对于监测这些移植细胞的命运以及评估其移植的安全性和有效性将具有重要价值。