PDF(Pubmed)
Abstract：
Human retinal organoids (ROs) have emerged as valuable tools for studying retinal development, modeling human retinal diseases, and screening drugs. However, their application is limited primarily due to time-intensive generation, high costs, and low reproducibility. Quality assessment of RO differentiation is crucial for their application in research. However, traditional methods such as morphological evaluation and immunohistochemical analysis have limitations due to their lack of precision and invasiveness, respectively. This study aims to identify non-invasive biomarkers for RO differentiation quality using exosomal microRNAs (miRNAs), which are known to reflect cell-specific functions and development in the retina. We differentiated ROs from human induced pluripotent stem cells (hiPSCs) and classified them into \'superior\' and \'inferior\' groups based on morphological and immunohistochemical criteria. Exosomes from the conditioned media were isolated and analyzed for miRNA content. Our findings revealed distinct miRNA profiles between superior and inferior ROs, with superior ROs exhibiting higher miRNA diversity and specifically up- or down-regulated miRNAs. Gene ontology and pathway enrichment analyses indicated that the target genes of these miRNAs are involved in neuron proliferation and differentiation. The study suggests the potential of exosomal hsa-miR-654-3p and hsa-miR-451a as non-invasive biomarkers for real-time monitoring of RO quality, facilitating the development of standardized, efficient, and cost-effective culture methods.
摘要：
人类视网膜类器官（RO）已经成为研究视网膜发育的有价值的工具，模拟人类视网膜疾病，和筛选药物。然而,它们的应用受到限制，主要是由于时间密集的生成，高成本,和低重现性。RO分化的质量评估对于其在研究中的应用至关重要。然而,形态学评价和免疫组织化学分析等传统方法由于缺乏精确性和侵入性而存在局限性，分别。本研究旨在使用外泌体microRNAs（miRNA）鉴定RO分化质量的非侵入性生物标志物，已知反映细胞特异性功能和视网膜发育。我们从人类诱导多能干细胞（hiPSCs）中分化出RO，并根据形态学和免疫组织化学标准将它们分为“上”和“下”组。分离来自条件培养基的外来体并分析miRNA含量。我们的发现揭示了不同的miRNA在上级和下级RO之间，具有表现出较高miRNA多样性的优异ROs，特别是上调或下调的miRNA。基因本体论和通路富集分析表明，这些miRNA的靶基因参与神经元的增殖和分化。该研究表明，外泌体hsa-miR-654-3p和hsa-miR-451a作为实时监测RO质量的非侵入性生物标志物的潜力。促进标准化的发展，高效,和具有成本效益的培养方法。
