Li Na, Li Xiao-rong, Yuan Jia-qin
Eye Center, Tianjin Medical University, Tianjin, 300070, China.
Graefes Arch Clin Exp Ophthalmol. 2009 Apr;247(4):503-14. doi: 10.1007/s00417-008-1009-y. Epub 2008 Dec 16.
To examine the survival, migration, integration, differentiation and the expression of various neurotrophic factors of bone-marrow mesenchymal stem cells (BMSCs) transplanted into the vitreous cavity of rats injured by ischemia/reperfusion(I/R).
The BMSCs were separated from rat marrow using the wall-sticking method, and cultured in vitro to expand. Flow cytometry detected the surface antigens of BMSCs. Ninety-six rats were randomly divided into four groups: normal control injected PBS(C+P), normal control injected BMSCs (C+B), ischemic/reperfusion injected PBS(I/R+P)and ischemic/reperfusion injected BMSCs(I/R+B). After retinal I/R injury was induced in each group by increasing intraocular pressure, 10 microl PBS and BMSC suspensions labeled by red fluorescence CM-Dil were immediately injected into the vitreous cavity. We observed the survival, migration and integration of BMSCs using confocal microscopy. The differentiation and expression of basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) of CM-Dil-labeled BMSCs were detected by immunofluorescent labeling and reserved by confocal microscopy. The expression of mRNA and proteins of bFGF, BDNF and CNTF were assayed by RT-PCR and Western Blot respectively.
After transplantation to normal eyes, BMSCs labeled by CM-Dil were mostly present in the vitreous cavity, and did not migrate. After transplantation to I/R eyes, BMSCs labeled by CM-Dil were mostly present along with the inner limiting membrane. Only a few cells were integrated into the ganglion cell layer. Two or 4 weeks after transplantation, a few BMSCs labeled by CM-Dil were observed to express markers of neuron- neurone specific enolase (NSE), neurofilament (NF) and various neurotrophic factors. The BMSC-injected I/R model eyes showed less reduction in the number of RGCs than that of the I/R eyes with PBS injection.
BMSC transplantation is a valuable neuroprotection tool for the treatment of retina and optic nerve diseases.
研究骨髓间充质干细胞（BMSCs）移植到缺血/再灌注（I/R）损伤大鼠玻璃体腔后的存活、迁移、整合、分化以及各种神经营养因子的表达情况。
采用贴壁法从大鼠骨髓中分离BMSCs，并进行体外培养扩增。流式细胞术检测BMSCs的表面抗原。96只大鼠随机分为四组：正常对照组注射PBS（C+P）、正常对照组注射BMSCs（C+B）、缺血/再灌注组注射PBS（I/R+P）和缺血/再灌注组注射BMSCs（I/R+B）。通过升高眼压诱导每组大鼠视网膜I/R损伤后，立即将10微升PBS和用红色荧光CM-Dil标记的BMSC悬液注入玻璃体腔。利用共聚焦显微镜观察BMSCs的存活、迁移和整合情况。通过免疫荧光标记检测CM-Dil标记的BMSCs中碱性成纤维细胞生长因子（bFGF）、脑源性神经营养因子（BDNF）和睫状神经营养因子（CNTF）的分化和表达，并通过共聚焦显微镜保存。分别采用RT-PCR和Western Blot检测bFGF、BDNF和CNTF的mRNA和蛋白表达。
移植到正常眼后，CM-Dil标记的BMSCs大多存在于玻璃体腔中，未发生迁移。移植到I/R眼后，CM-Dil标记的BMSCs大多与内界膜一起存在。仅有少数细胞整合到神经节细胞层。移植后2周或4周，观察到少数CM-Dil标记的BMSCs表达神经元特异性烯醇化酶（NSE）、神经丝（NF）等神经元标志物以及各种神经营养因子。注射BMSCs的I/R模型眼视网膜神经节细胞（RGCs）数量的减少程度低于注射PBS的I/R眼。
BMSC移植是治疗视网膜和视神经疾病的一种有价值的神经保护工具。
