Abstract
Abstract Purpose The experimental data on the retinal pigment epithelium (RPE) transplantation clearly show the potential of the cell therapy treatment strategy for RPE dysfunction. Human embryonic stem cells (hESC) may have a significant role as tools for drug discovery and for replacement of damaged cells. Human ESCs are potent origin for mass production of specific cells, thus providing unlimited opportunities for these strategies. However, the utilization of hESC‐derived RPE cells in therapy requires an efficient and xeno‐free differentiation protocol that follows good manufacturing practice (GMP) standards. In addition, it is essential to develop methods for reliable characterization of the cells to gain safe and clinically eligible cells. Methods We have compared the differentiation potential of several hESC lines and induced RPE differentiation in the absence of animal feeder cells and bovine serum. The differentiation of cells has been followed by expression analysis of retinal cell markers by using qRT‐PCR and protein analysis. Results With our differentiation protocol, we were able to generate RPE cells from several hESC lines with satisfying efficiency. The typical pigmented cobblestone‐like morphology and expression of retinal precursor markers were detected within one week. Further, RPE specific markers were up‐regulated during the maturation of the cells. Conclusion We have developed progressive differentiation protocol, which enables generation of RPE cells without animal feeder cells and serum induction. The generated cell material is of high quality and more compatible with GMP requirements than cells generated in previous studies. In my presentation, I will review some of the data using hESC for RPE production and our experience in utilizing the hESC‐derived RPE cells.
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