Department of Pharmacy, University of Pisa, Via Bonanno Pisano, 6, 56126, Pisa, Italy.
Scuola Superiore Sant'Anna, Pisa, Italy.
Sci Rep. 2023 Apr 13;13(1):6025. doi: 10.1038/s41598-023-32571-y.
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.
在增殖的多能视网膜祖细胞中，转录因子的动态变化决定了有丝分裂后子细胞的命运，但由外在因素驱动的有丝分裂后细胞命运可塑性仍存在争议。转录组分析揭示了有丝分裂后杆状前体细胞同时表达对 Müller 胶质细胞命运至关重要的基因，而这些基因很少与杆状前体细胞一起由终末分裂的祖细胞产生。通过在单个培养的杆状前体细胞中结合基因表达和功能特征，我们确定了一个时间限制的窗口，在此期间，增加细胞培养密度会关闭对 Müller 胶质细胞命运至关重要的基因的表达。有趣的是，在低细胞培养密度下的杆状前体细胞保持杆状和神经胶质细胞命运的基因表达，并表现出混合的杆状/Müller 胶质细胞电生理特征，表明杆状细胞向混合的杆状-神经胶质表型偏离。细胞培养密度作为一个关键的外在因素，可以防止杆状命运的细胞向混合细胞状态偏离的观点，可能解释了在成年视网膜中混合杆状/MG 细胞的发生，并为通过稳定移植的杆状前体细胞的命运来改善再生方法治疗视网膜退行性疾病的移植物植入率提供了一种策略。
