RESUMO
With recent advances in adeno-associated virus (AAV)-based gene therapy, efficacy and toxicity screening have become essential for developing gene therapeutic drugs for retinal diseases. Retinal organoids from human pluripotent stem cells (hPSCs) offer a more accessible and reproducible human test platform for evaluating AAV-based gene therapy. In this study, hPSCs were differentiated into retinal organoids composed of various types of retinal cells. The transduction efficiencies of AAV2 and AAV8, which are widely used in clinical trials of inherited retinal diseases, were analyzed using retinal organoids. These results suggest that retinal organoids derived from hPSCs serve as suitable screening platforms owing to their diverse retinal cell types and similarity to the human retina. In summary, we propose an optimal stepwise protocol that includes the generation of retinal organoids and analysis of AAV transduction efficacy, providing a comprehensive approach for evaluating AAV-based gene therapy for retinal diseases.
RESUMO
Given the advantage of being able to be extracted by a minimally invasive method, blood is regarded as a suitable cell source for reprogramming to establish induced pluripotent stem cells (iPSCs). Therefore, iPSCs established from patient derived peripheral blood mononuclear cells (PBMCs) is widely used to develop disease modeling to elucidate disease development. Here, PBMCs from a healthy man were reprogrammed into iPSCs using the Sendai virus. The established iPSC line, KRIBBi006-A, exhibit pluripotency marker and can differentiate into the three germ layers in vitro with normal karyotype. This iPSC line is a valuable resource as a control line for stem cell research of disease models and drug screening.
