Bone marrow stromal cell transplantation as the treatment for glaucomatous optic neuropathy.

为了了解骨髓基质细胞（BMSCs）移植是否有利于青光眼的重建或保护，我们研究了移植的骨髓基质细胞在青光眼模型眼中是否能够存活、分化为视网膜神经节细胞（RGCs），并防止宿主青光眼视网膜RGC的减少。结扎骨膜外静脉诱导成年大鼠右眼青光眼，假手术左眼作为对照。将表达gfp的骨髓间充质干细胞（GFP-BMSCs）或磷酸盐缓冲盐水（PBS）注射到对照组和不同阶段青光眼的玻璃体中。移植1周后，GFP-BMSCs多与内限制膜（ILM）一起存在，少数细胞融入神经节细胞层（GCL）。视盘内未见细胞整合。移植细胞表达间充质细胞标志物CD54，但未观察到神经元或胶质细胞标志物的表达。注射后4周氟金逆行标记RGC数量分析显示，与PBS注射青光眼相比，BMSC注射青光眼的RGC数量减少较少。移植后2、4周，免疫组化观察GFP-BMSCs表达多种营养因子。实时荧光定量PCR检测结果显示，BMSC移植青光眼中bFGF和CNTF的表达明显高于PBS注射青光眼。综上所述，BMSCs在青光眼中能够存活并表达营养因子，且BMSCs移植青光眼视网膜可阻止RGC数量的减少。骨髓间充质干细胞移植可能是治疗青光眼的一种神经保护工具。

To know if the transplantation of bone marrow stromal cells(BMSCs) could be beneficial for reconstructing or protecting glaucomatous eyes, we investigated if transplanted BMSCs in glaucoma model eye could survive, differentiate into retinal ganglion cells(RGCs), and also prevent the reduction of RGC in host glaucoma retina. Glaucoma was induced in the right eye of adult rat by ligating episcleral veins and the left eye was sham operated and used as a control. The GFP-expresssing BMSCs(GFP-BMSCs) or phosphate buffered saline(PBS) were injected into the vitreous body of both the control and the glaucoma eyes at various stages. One week after transplantation, GFP-BMSCs were mostly present along with inner limiting membrane(ILM) and a few cells were integrated into the ganglion cell layer(GCL). No cells were integrated into the optic disc. The transplanted cells expressed CD54, the marker of mesenchymal cells, however the expression of neuronal or glial markers were not observed. The analysis of RGC number by fluorogold retrogradely labeling at 4 weeks after injection revealed less reduction in the number of RGCs in BMSC injected glaucoma eyes compared with the PBS injected glaucoma eyes. At 2 or 4 weeks after transplantation, GFP-BMSCs were observed to express various trophic factors by immunohistochemistry. The quantative assessment by real-time PCR demonstrated that the expression of of bFGF and CNTF in BMSC transplanted glaucoma eyes is significantly increased compared with the PBS injected glaucoma eyes. In conclusion, BMSCs could survive and express trophic factors in glaucoma eyes and the decrease in the number of RGC was prevented in BMSCs transplanted glaucoma retina. BMSCs transplantation may be worthy as a neuroportective tool to treat glaucoma.