Graduate Student Chungbuk National University cheongju, Republic of Korea
Abstract: Porcine cerebral organoids have multiple important applications in neuroscience research and drug development. Derived from pluripotent stem cells, human cerebral organoids can self-organize into three-dimensional structures that replicate key features of the human brain, such as cortical layers and neurogenesis. This capability allows them to be effectively used to model various neurological disorders. Given the ethical challenges associated with using human tissue for invasive brain development studies, the development of porcine models presents a more effective alternative for investigating disease mechanisms. However, traditional in vivo models, such as transgenic pigs, can be time-consuming and costly to produce. Therefore, the establishment of in vitro models like porcine cerebral organoids may offer a more efficient, ethical, and cost-effective approach to studying human neurobiology and pathology. In this study, by using in vitro fertilization and somatic cell nuclear transfer-derived porcine embryonic stem cells, we primarily used an adapted Lancaster Human Cerebral Organoid Generation Protocol to better suit the development of porcine cerebral organoids. Early porcine cerebral organoids exhibited morphological changes similar to those observed in early human cerebral organoids under bright fields. Immunofluorescence analysis of porcine cerebral organoids confirmed that the organoids were undergoing neural differentiation. At early development , the porcine brain organoids showed strong expression of neural progenitor markers, including PAX6, NESTIN, SOX1, and ZO1. At the midpoint of development, the porcine cerebral organoid exhibited signals indicative of neural progenitor regions and early neurogenesis. They displayed the formation of ventricular-like cavities, with SOX2 (inner layer) and TUJ1(outer layer) staining. By day 50, the porcine cerebral organoid showed positive expression of GFAP around areas expressing NESTIN, and IBA1 showed weak positivity at the margins of the organoid. PAX6 expression was minimal. In conclusion, the establishment of porcine brain organoid models offers an additional and valuable in vitro tool for studying neurological disorders.
Funding Source: This work was granted by the “Technology Innovation Program funded by the MOTIE, Korea (20023068)”, and “IPET in Food, Agriculture, Forestry and Fisheries funded by MAFRA (RS-2024-00398561, RS-2024-00399475)", Republic of Korea.
