Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical University, 1-25-4 Daigaku-nishi, Gifu 501-1196, Japan.
Exp Eye Res. 2013 Nov;116:254-64. doi: 10.1016/j.exer.2013.09.013. Epub 2013 Sep 26.
Human adipose-derived stem cells (hASCs) are present in adult adipose tissue and have been reported to secrete various factors that have neuroprotective effects. In the present study, we examined whether hASC-conditioned medium (hASC-CM) was effective against experimental degenerative retinal disease. Mature adipocytes (MAs) and hASCs were isolated from human subcutaneous adipose tissue. The isolated hASCs were identified based on their capacity for bone and neural differentiation. The effects of hASC-CM against tunicamycin-, H2O2-, and light-induced retinal photoreceptor damage were evaluated in vitro by measuring cell death. Moreover, we identified various factors present in hASC-CM using antibody arrays. Retinal damage induced in mice by exposure to white light was studied in vivo, and photoreceptor damage was evaluated according to the thickness of the outer nuclear layer and electroretinography results. In addition, the effect of hASC-CM on Akt phosphorylation at Ser473 was confirmed by western blotting. Finally, the effects of the secreted proteins identified in the hASC-CM on light-induced damage were evaluated in vivo. Isolated hASCs differentiated to osteocytes and neurons. hASC-CM protected against tunicamycin-, H2O2-, and light-induced cell death. In addition, hASC-CM inhibited photoreceptor degeneration and retinal dysfunction after exposure to light. Several proteins secreted by hASCs, such as the tissue inhibitor of metalloproteinase-1 (TIMP-1) and the secreted protein acidic and rich in cysteine (SPARC), protected against light-induced damage in vitro and in vivo. The results of the present study showed that hASC-CM has neuroprotective effects against light-induced retinal damage and suggest that hASCs have a therapeutic potential in retinal degenerative diseases via their secreted proteins, without requiring transplantation.
人脂肪来源的干细胞（hASCs）存在于成人脂肪组织中，并已被报道分泌具有神经保护作用的各种因子。在本研究中，我们研究了 hASC 条件培养基（hASC-CM）是否对实验性退行性视网膜疾病有效。从人皮下脂肪组织中分离成熟脂肪细胞（MAs）和 hASCs。通过骨和神经分化能力鉴定分离的 hASCs。通过测量细胞死亡来评估 hASC-CM 对衣霉素、H2O2 和光照诱导的视网膜光感受器损伤的体外作用。此外，我们使用抗体阵列鉴定了 hASC-CM 中存在的各种因子。通过暴露于白光在体内研究了诱导的小鼠视网膜损伤，并根据外核层厚度和视网膜电图结果评估光感受器损伤。此外，通过 Western blot 验证了 hASC-CM 对 Akt 丝氨酸 473 磷酸化的影响。最后，评估了 hASC-CM 中鉴定的分泌蛋白对光诱导损伤的体内作用。分离的 hASCs 分化为成骨细胞和神经元。hASC-CM 可防止衣霉素、H2O2 和光照诱导的细胞死亡。此外，hASC-CM 可抑制光照后的光感受器变性和视网膜功能障碍。hASCs 分泌的几种蛋白质，如基质金属蛋白酶组织抑制剂-1（TIMP-1）和富含半胱氨酸的酸性分泌蛋白（SPARC），在体外和体内均能抵抗光诱导的损伤。本研究结果表明，hASC-CM 对光诱导的视网膜损伤具有神经保护作用，并提示 hASCs 通过其分泌蛋白在视网膜退行性疾病中具有治疗潜力，而无需移植。
