Gene expression profile of cells from cochlear organoids and comparison with in vivo auditory sensory epithelium cells.

We sequenced the total RNA of cells from inner ear organoids generated by EFI,EFI-4C and EFI-CL expansion strategies. Transcriptional profiles revealed that CHIR and LPA significantly increased proliferative potency and contributed to the stemness maintenance. We further compared the gene expression profiles among the sorted Lgr5+ CPCs, the primary organoids of passage (P)0, the P4 organoids, and the sorted E-cadherin+ sensory epithelial cells. Inner ear organoids recapitulated genetic characteristics of auditory epithelium rather than Lgr5+ progenitors alone. Differential gene analysis of cells from primary organoids and organoids at fourth passage indicated stable transcriptional profiles among passages, which confirmed the reliability and authenticity of cochlear organoids for modeling the properties of the cochlea. Transcriptional profile of inner ear organoids generated by different expansion strategies, Lgr5+ CPCs and E-cadherin+ sensory epithelial cells.

本研究对采用EFI、EFI-4C及EFI-CL三种扩增策略构建的内耳类器官（inner ear organoids）中的细胞总RNA进行了测序。转录组分析结果显示，CHIR与LPA可显著提升细胞增殖能力，并有助于维持干细胞干性（stemness）。本研究进一步比较了分选得到的Lgr5阳性祖细胞（Lgr5+ CPCs）、第0代（P0）原代类器官、第4代（P4）类器官，以及分选得到的E-钙粘蛋白阳性（E-cadherin+）感觉上皮细胞的基因表达谱。内耳类器官可重现听觉上皮的遗传特征，而非仅复刻Lgr5阳性祖细胞的特性。对原代类器官与第4代类器官的细胞进行差异基因分析后发现，不同代次的类器官转录组谱保持稳定，这证实了耳蜗类器官（cochlear organoids）用于模拟耳蜗特性的可靠性与真实性。本数据集涵盖采用不同扩增策略构建的内耳类器官、Lgr5阳性祖细胞及E-钙粘蛋白阳性感觉上皮细胞的转录组谱数据。