绿色荧光蛋白转基因小鼠CD34^+/CD45^+细胞在脊髓全横断大鼠模型中的存活及迁移

Survival and distribution of CD34^+/CD45^+ cells from bone marrow of transgenic GFP mouse in a completely transected spinal cord rat model

目的:探讨CD34+/CD45+细胞移植入脊髓全横断大鼠模型后的存活、迁移及分布情况。方法:体外培养绿色荧光蛋白(GFP)转基因小鼠骨髓细胞,经CD34、CD45单克隆抗体鉴定后移植入脊髓全横断大鼠模型脊髓横断处尾侧,分别在术后24h、48h、1周、2周、4周和8周行左心腔内灌注,取出脊髓,连续切片(片厚10μm),置于荧光显微镜下观察切片中有无绿色荧光细胞,并观察荧光细胞的分布范围;用免疫组化法检测CD34+/CD45+细胞的存活。结果:脊髓横断处头尾两侧均可见绿色荧光细胞,且多分布于灰质中,散在或聚集成片;免疫组化法可见切片中有CD34+/CD45+细胞散在。结论:CD34+/CD45+细胞可在脊髓全横断大鼠模型的脊髓中存活,并可迁移至脊髓横断处头侧,且随时间的延长迁移距离有所增加。

AIM： To study the survival, transfer and distribution of bone marrow CD34 ^＋/CD45^＋ cells from transgenic GFP mouse after transplanted into the completed transversional spinal cord rat model. METHODS： The bone marrow cells isolated from transgenic GFP mice were cultured in vitro. The cultured cells were identified by anti - CD34 and anti - CD45 monoclonal antibodies, and were transferred into the end of transection spinal cord. Paraformaldehyde was infused into the left ventricle of the rat model at the 24 h, 48 h, 1 week, 2 weeks, 4 weeks and 8 weeks after cell transplantation. Through sank and frozen, the spinal cord was sectioned at 10 um thickness. The green fluorescence positive cells were observed under the fluorescence microscope. CD34 ^＋/CD45^＋ cells were identified by immunohistochemistry staining. RESULTS： Green fluorescence positive cells were found at the head and the end of the completed transection part of spinal cord. Most of the green fluorescence positive cells were distributed in the gray substance of spinal cord. CD34 ^＋/ CD45 ^＋ cells were found by immunohistochemistry staining. CONCLUSION： CD34^＋/CD45^＋ cells survived in spinal cord of SD rat, and migrated to the head of the transection part. The distance of migration was extended by the time.