Medical Physics and Biomedical Engineering Department, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Neurosci Lett. 2013 Apr 29;541:43-8. doi: 10.1016/j.neulet.2012.12.055. Epub 2013 Feb 8.
Stem cell therapy is a promising approach for treatment of degenerative retinal disorders such as retinitis pigmentosa (RP) and age-related macular degeneration (AMD). In this study, human mesenchymal stem cells (MSCs) were isolated from the trabecular meshwork (TM), the major functional tissue of the anterior chamber angle in the eye, were characterized and differentiated into photoreceptor cells on amniotic membrane (AM). After isolation of trabecular meshwork and culture of the stromal segment of this tissue, fibroblast-like cells (CD105(+), CD90(+), CD44(+), CD166(+) cells) capable of differentiation toward mesenchymal and photoreceptor lineages were obtained. The isolated cells were seeded on amniotic membrane and were treated with induction medium. Immunocytochemistry and quantitative real time RT-PCR (qPCR) were used to detect expression of photoreceptor genes such as rhodopsin, recoverin, CRX, and peripherin; and the bipolar cell marker protein kinase C alpha (PKC-alpha). As a result, immunocytochemistry revealed that the differentiated TMMSCs expressed rhodopsin, CRX and PKC proteins. qPCR showed the expression of rhodopsin (rod like photoreceptor-specific marker), and CRX genes were significantly higher in TMMSCs differentiated on AM than those differentiated on tissue culture polystyrene (TCPS). In conclusion, our findings suggested that a combination of TMMSCs (as a new source) and basement membrane support from AM might be a suitable source of cells for subretinal transplantation in regenerative therapy for retinal disorders such as AMD and RP.
干细胞疗法是治疗退行性视网膜疾病（如色素性视网膜炎（RP）和年龄相关性黄斑变性（AMD））的一种很有前途的方法。在这项研究中，从小梁网（TM）中分离出人骨髓间充质干细胞（MSCs），TM 是眼睛前房角的主要功能组织，在羊膜上分化为光感受器细胞。在分离小梁网并培养该组织的基质段后，获得了能够向间充质和光感受器谱系分化的成纤维样细胞（CD105（+）、CD90（+）、CD44（+）、CD166（+）细胞）。分离的细胞被接种到羊膜上，并使用诱导培养基进行处理。免疫细胞化学和实时定量 RT-PCR（qPCR）用于检测光感受器基因如视蛋白、恢复蛋白、CRX 和周围蛋白的表达；和双极细胞标记蛋白激酶 C alpha（PKC-alpha）。结果表明，免疫细胞化学显示分化的 TMMSCs 表达视蛋白、CRX 和 PKC 蛋白。qPCR 显示视蛋白（杆状光感受器特异性标记物）的表达，并且在 AM 上分化的 TMMSCs 比在组织培养聚苯乙烯（TCPS）上分化的 TMMSCs 中 CRX 基因的表达显著更高。总之，我们的研究结果表明，TMMSCs（作为一种新来源）与 AM 基底膜支持的结合可能是用于 AMD 和 RP 等视网膜疾病的视网膜下移植再生治疗的合适细胞来源。
