Human Adipose-derived Stem Cells Cultured In Modified Conditions Show Improved Protective Functions To Retinal Neurons

Objective:To investigate the effects of multipotent adult progenitor cell (MAPC) culture conditions on biological characteristics and therapeutic effects of human adipose-derived stem cells (hADSCs), thus to obtain optimal donor cells for treatment of retinal degeneration (RD).Methods:The cell morphology, MTT assay, clone formation, adipogenic, osteogenic, chondrogenic differentiation potential and qRT-PCR technique were performed to detect biological characteristics of hADSCs. The hADSCs were cultured under MAPC culture conditions and transplanted into the subretinal space of RCS rats. The retinal protective functions were evaluated by electroretingram (ERG) recording, histological examination and TUNEL assay.Results:While MAPC culture conditions, compared with conventional hADSCs culture medium, maintained hADSCs morphology in long-term culture (beyond10passages), and the cells displayed lower senescence ratio, higher proliferative capacity and multi-lineage differentiation potentials. Gene expression profiles for cell surface markers and cytokines showed a differential expression patterns:Under MAPC culture conditions, CD140b, CD90, CD47, HGF and PEDF was significantly up-regulated, whereas CD73, CD105and IL-6were significantly down-regulated compared with those under conventional culture medium. In comparison with untreated RCS rats, MAPC-hADSCs transplanted RCS rats showed significantly improved b-wave amplitudes in ERG examination in3weeks after the transplantation, and significant reduction of apoptotic cells in the outer nuclear layer (ONL).Conclusions:hADSCs treated with MAPC culture conditions showed improved retinal neuro-protection and hold great promise for clinical use in RD disease.