Differenzierung humaner IPSC-angeleiteter optisch neuronaler Vorläuferzellen für die Behandlung der auditorischen Neuropathie

In the past decade, harnessing the potential of progenitor cells for regeneration of the auditory nerve evolved as a new approach to the treatment of cochlear neuropathy e.g. the loss of spiral ganglion neurons (SGN). Learning that CI performance is correlated with the number of healthy SGN in the cochlea has prompted the hypothesis that replacing SGNs could enhance hearing restoration in this patient population. This study explores the potential of optogenetically modified otic neuronal progenitor cells (ONP) derived from human induced pluripotent stem cells (iPSCs) to build SGN-like cells for regeneration of the auditory nerve. We generated otic bioengineered neural organoids (oBENO) by patterning human iPSC towards an otic fate by a stage-specific differentiation similar to in vivo development. Parvalbumin-positive ONP can be traced due to an integrated EGFP-tag in the PVALB-gene. For the enhancement of ONP differentiation, the effect of adding neuronal factors was analyzed. The growth factors BDNF (Brain Derived Neurotrophic Factor), NT-3 (Neurotrophin-3) GDNF (Glial Cell line-Derived Neurotrophic Factor) and NGF (Nerve Growth Factor) and combinations of factors were added to the media of whole oBENOs and digested neurons in 2D-culture in hydrogel starting from day 20 to day 40 every two days. Cell differentiation and neuronal growth were examined by live-cell imaging and immunofluorescence staining. Preliminary data show, that administration of a single growth factor already exerts a positive effect on cell differentiation as well as survival. However, the combination of all three factors, BDNF, NT-3 and NGF, has demonstrated an even more pronounced effect. This may provide a foundation for future research on auditory nerve regeneration in cases of SGN loss.

Nein