Scientific Reports (Apr 2023)

Increasing cell culture density during a developmental window prevents fated rod precursors derailment toward hybrid rod-glia cells

  • Ivana Barravecchia,
  • Chiara De Cesari,
  • Viviana Guadagni,
  • Giovanni Signore,
  • Edoardo Bertolini,
  • Serena Gea Giannelli,
  • Francesca Scebba,
  • Davide Martini,
  • Mario Enrico Pè,
  • Vania Broccoli,
  • Massimiliano Andreazzoli,
  • Debora Angeloni,
  • Gian Carlo Demontis

DOI
https://doi.org/10.1038/s41598-023-32571-y
Journal volume & issue
Vol. 13, no. 1
pp. 1 – 17

Abstract

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Abstract In proliferating multipotent retinal progenitors, transcription factors dynamics set the fate of postmitotic daughter cells, but postmitotic cell fate plasticity driven by extrinsic factors remains controversial. Transcriptome analysis reveals the concurrent expression by postmitotic rod precursors of genes critical for the Müller glia cell fate, which are rarely generated from terminally-dividing progenitors as a pair with rod precursors. By combining gene expression and functional characterisation in single cultured rod precursors, we identified a time-restricted window where increasing cell culture density switches off the expression of genes critical for Müller glial cells. Intriguingly, rod precursors in low cell culture density maintain the expression of genes of rod and glial cell fate and develop a mixed rod/Muller glial cells electrophysiological fingerprint, revealing rods derailment toward a hybrid rod-glial phenotype. The notion of cell culture density as an extrinsic factor critical for preventing rod-fated cells diversion toward a hybrid cell state may explain the occurrence of hybrid rod/MG cells in the adult retina and provide a strategy to improve engraftment yield in regenerative approaches to retinal degenerative disease by stabilising the fate of grafted rod precursors.