Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid, Valladolid, Spain.
Histol Histopathol. 2013 Dec;28(12):1577-83. doi: 10.14670/HH-28.1577. Epub 2013 May 30.
Age-related macular degeneration is a retinal disease with important damage at the RPE layer. This layer is considered a target for therapeutical approaches. Stem cell transplantation is a promising option for retinal diseases. Adipose derived mesenchymal stem cells secret growth factors which might play a significant role in RPE maintenance. This study aimed to evaluate human AD-MSCs ability to rescue mitomycin C treated dying ARPE19 cells in co-culture condition. ARPE19 cells were treated with MMC (50 μg/ml, 100 μg/ml and 200 μg/ml) for 2 hours to induce cell death. These treated cells were co-cultured with hAD-MSCs in indirect co-culture system for 3 days and 3 weeks. Then the viability, growth and proliferation of these ARPE19 cells were evaluated by a cell viability/cytotoxicity assay kit and Alamar Blue (AB) assay. Untreated ARPE19 cells and human skin fibroblasts (HSF) were used as controls. MMC blocked ARPE19 cell proliferation significantly in 3 days and cells were almost completely dead after 3 weeks. Cell toxicity of MMC increased significantly with concentration. When these cells were co-cultured with hAD-MSCs, a significant growth difference was observed in treated cells compared to untreated cells. hAD-MSCs rescue capacity was also significantly higher than HSF for treated ARPE19 cells. This study showed that hAD-MSCs rescued MMC treated ARPE19 cells from death. It probably occurred due to undefined growth factors secreted by hAD-MSCs in the medium, shared by treated ARPE19 cells in co-culture conditions. This study supports further evaluation of the effect of hAD-MSCs subretinal transplantation over the RPE degeneration process in AMD patients.
年龄相关性黄斑变性是一种视网膜疾病,在 RPE 层有重要损伤。该层被认为是治疗方法的靶点。干细胞移植是治疗视网膜疾病的一种有前途的选择。脂肪来源的间充质干细胞分泌的生长因子可能在 RPE 维持中发挥重要作用。本研究旨在评估人 AD-MSCs 在共培养条件下拯救丝裂霉素 C 处理的 ARPE19 细胞死亡的能力。ARPE19 细胞用 MMC(50μg/ml、100μg/ml 和 200μg/ml)处理 2 小时以诱导细胞死亡。这些处理过的细胞与 hAD-MSCs 在间接共培养系统中共同培养 3 天和 3 周。然后通过细胞活力/细胞毒性测定试剂盒和 Alamar Blue(AB)测定评估这些 ARPE19 细胞的活力、生长和增殖。未处理的 ARPE19 细胞和人皮肤成纤维细胞(HSF)用作对照。MMC 在 3 天内显著阻断 ARPE19 细胞的增殖,并且在 3 周后细胞几乎完全死亡。MMC 的细胞毒性随浓度显著增加。当这些细胞与 hAD-MSCs 共培养时,与未处理的细胞相比,处理过的细胞观察到明显的生长差异。与处理过的 ARPE19 细胞相比,hAD-MSCs 对 HSF 的拯救能力也明显更高。本研究表明,hAD-MSCs 从死亡中拯救了 MMC 处理的 ARPE19 细胞。这可能是由于 hAD-MSCs 在培养基中分泌的未定义生长因子在共培养条件下被处理的 ARPE19 细胞共享所致。本研究支持进一步评估 hAD-MSCs 视网膜下移植对 AMD 患者 RPE 变性过程的影响。