Compositions and methods for efficient amplification of retinal progenitors cells
Human induced pluripotent stem (hiPS) cells can be used as an unlimited source of retinal cells for the treatment of retinal degenerative diseases. Moreover, different field of application establishes potential roles for iPSCs in modeling diseases and drug screening.
Although much progress has been made in the differentiation of pluripotent stem cells towards different retinal lineages, classical techniques do not enable the production of a huge amount of retinal cells from hiPSC. Indeed, cultured cells often left multipotency and/or their amplification properties. This requires the development of easy and standardized protocols to obtain billion cells.
To address these requirements, Dr. Olivier Goureau’s team, at the Vision Institute, developed a new culture medium allowing simple and effective amplification and differentiation of retinal progenitor cells (RPC) obtained from hiPSC.
This medium induce the generation of the main retinal cell types: retinal ganglion cells, precursors of photoreceptor and Müller cells. Those differentiated cells are obtained in a huge quantity (billion of cells) and are functionals. Cells can even be applied for therapeutic purposes because the same phenotypes are obtained and results are reproducible,
• The ability of RPC to differentiate in PRs and RGCs by spontaneous differentiation was tested. The results obtained by immunostaining show that the RPC at passage 2 to 6 are able to differentiate in the photoreceptor precursor and retinal ganglion cell ways.
• The passage number was determined by performing a series of consecutive passages each week. The results show that the cells cultured and passed several times are still able to multiply up to the RPCp7 stage.
• The multipotent and mitotic character was confirmed by a RT-qPCR analysis. Results show that from the stage RPCp2 to RPCp4, the expression of RPC-specific genes remains stable and comparable to native RPCs.