Recently, we are exploring the use of human corneal organoids as an animal alternative model system to investigate human corneal development and disease. Matured for up to 4 months in culture, these human corneal organoids establish a layered ECM-rich cornea-like tissue (Foster, 2017), and harbor cells that mimic all three major cell types of the cornea and a rudimentary layered ECM with collagen fibrils (Maiti, 2022). We are exploring the human corneal organoids as a model system for 1) corneal repair and regeneration after injury or infections, 2) as a source of ECM and stromal cells for corneal therapy, and 3) understanding functions of novel genes in the cornea. Here we are investigating cell and ECM regeneration in the corneal organoids using transcriptomic, RNAscope, immunohistology, confocal and electron microscopy.