1. Development of Non-Invasive miRNA Markers for Assessing the Quality of Human Induced Pluripotent Stem Cell-Derived Retinal Organoids
- Author
-
Hyo Song Park, Ji-Hong Bang, Wook Hyun Jung, Jin Young Yang, Hee Jeong Shin, Ji-Hye Son, Jung Woo Han, Si Hyung Lee, Kyung Hwun Chung, Kyunggon Kim, Hun Soo Chang, and Tae Kwann Park
- Subjects
retinal organoids ,exosomes ,microRNA ,differentiation ,non-invasive ,markers ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - 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.
- Published
- 2024
- Full Text
- View/download PDF