Molekulare und funktionelle Charakterisierung von Stamm- und Progenitorzellen der menschlichen Hornhaut
Zusammenfassung der Projektergebnisse
Despite the difficulties to obtain primary limbal tissue after the relocation of Prof. Meller in 2015, we successfully reorganized the project. The intention was to establish culture conditions allowing for an expansion of limbal epithelial progenitor cells and to generate such cells de novo from pluripotent stem cells. Within the funding period, we were able to establish a) novel culture conditions allowing for extended growth of limbal epithelial cell progenitors, in vitro; b) a novel, inducible lentiviral H2B-GFP „all-in-one“ expression vector for the detection of label-retaining (progenitor) cells; c) an iPSCs diffentiation culture allowing for the development and enrichment of corneal epithelial cells, based on the protocol of Hayashi et al. (2016); d) enhanced generation of retinal pigment epithelial cells (RPE) from iPSCs ectopically expressing selected transcription factors involved in eye-development, particularly SIX2. Notably, this finding is of particular interest as iPSC-derived RPE cells are already used in a japanese phase I clinical trial for the treatment of age-related macular degeneration (AMD). Thus, methods which allow for an increased formation of patient-specific RPE, in vitro, may turn out to be highly relevant for future clinical application.
