Asan Medical Center, University of Ulsan College of Medicine, Republic of Korea
Abstract: Mitochondrial stress and dysfunction significantly impact cellular functions and contribute to the pathology of a variety of diseases, including Fuchs’ syndrome. Recent studies have explored the regulatory role of mitochondria derived from stem cells as a novel therapeutic approach for various diseases. We aim to investigate whether mitochondria isolated from induced pluripotent stem cell-derived corneal endothelial cells (iPSC-CEC) can restore the functionality of damaged CEC. Inflammatory conditions were induced in cells by treatment with lipopolysaccharide (LPS) and interferon-gamma (IFN-γ) to iPSC-CECs. We assessed the effects of iPSC-CEC mitochondria on mitochondrial morphology under inflammatory conditions, using confocal microscopy to observe mitochondrial rescue. We also evaluate the influence of mitochondrial delivery on cellular function and proliferation using an in vitro scratch assay and CCK8 assay. Finally, Western blot analysis was conducted to evaluate cell survival and the changes of inflammatory responses. Treatment with iPSC-CEC mitochondria successfully rescued mitochondrial fusion morphology under inflammatory conditions induced by LPS and IFN-γ, as shown by confocal microscopy. Delivery of mitochondria significantly reduced the expression of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α at both mRNA and protein levels. Scratch assays demonstrated increased cell migration and wound closure following mitochondrial treatment. Western blot analysis revealed increased Mfn-2 and p-AKT expression, indicating improved mitochondrial function and survival pathways. These results highlight the therapeutic efficacy of iPSC-CEC mitochondria in restoring cellular functions and reducing inflammation under pathological conditions of CECs, suggesting the potential of iPSC-CEC-derived mitochondria as a therapeutic agent in regenerative medicine of cornea.
Funding Source: This work was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (RS-2024-00438366)
