Towards adipose tissue-derived stromal cells-based therapy for diabetic retinopathy

In this thesis, we investigated the impact of hyperglycemia on adipose tissue-derived stromal cells (ASC) as a prelude to their use in the future therapeutic treatment of diabetic retinopathy (DR). Ultrastructure analyses of co-cultured ASC-endothelial cells featured the pericytic role of ASC in the maintenance of the vascular architecture under normal and high glucose conditions. The ROS-induced mitochondrial dysfunction and hyperglycemia-induced apoptosis partially influenced the pericytic functions of ASC. Injected ASC into an angiogenic mouse models, were detected at pericytic positions on newly formed vessels. Our recommendation is to preculturing of ASC under ‘chronic’ hyperglycemia, before injecting into a hyperglycemic environment to reduce their expression of pro-inflammatory and pro-angiogenic genes. ASC-conditioned medium (ASC-Cme) delivered from ASC cultured in chronically HG, protected bovine retinal endothelial cells (BREC) from hyperglycemia-induced apoptosis and inflammatory activation. This effect on BREC was evoked by the ROS-neutralizing capacity of ASC-Cme in culture that was associated with a reduced NF-κB activation which showed as a downmodulation of HG-upregulated pro-inflammatory genes in BREC. Towards using ASC for treatment of DR, we present a new hypothermic storing technique of cells in their own culture medium to well-maintain and transport the prepared cells to their clinical destination. The new pharmacologic compound, (6-hydroxyl-2,5,7,8-tetramethylchroman-2-yl)(4-(2-hydroxyethyl)piperazin-1-yl) methanone (SUL-109) shields ASC during cell preservation from hypothermic cell death without influencing their multi-potency capacity and proliferation through maintenance of the mitochondrial membrane potential and promoting the activation of mitochondrial complexes I and IV, consequently sustaining ATP production and preventing the overproduction of ROS under hypothermic conditions