Abstract Background Intestinal epithelial cells are supported by dynamic cellular heterogeneity, which is critical for maintaining intestinal homeostasis. Recently, intestinal organoid models have gained attention as in vitro platforms because they can recapitulate the structural, functional, and cellular complexity of the small intestine. In this study, we developed porcine intestinal organoids and investigated time-dependent transcriptomic changes using single-cell RNA sequencing. Furthermore, to assess their applicability as an in vitro toxicity model, the organoids were exposed to the mycotoxins deoxynivalenol and zearalenone. Results The established organoids exhibited stable long-term culture up to passage 10 (32 d) and showed high genetic similarity to native small intestinal tissue across three regions: the duodenum, jejunum, and ileum. In various intestinal epithelial cell types, including transit-amplifying cells, enteroendocrine cells, goblet cells, Paneth cells, and other epithelial cell types, were identified in the organoids. Single-cell RNA sequencing classified the organoids into multiple distinct cell populations, including stem cells, transit-amplifying cells, secretory progenitors, enterocytes, enteroendocrine cells, goblet cells, and Paneth cells, demonstrating dynamic cellular heterogeneity. The organoids also recapitulated key intestinal functions, such as nutrient absorption and epithelial barrier formation, similar to those of the native small intestinal epithelium. Under these conditions, the cytotoxic effects of deoxynivalenol and zearalenone were evaluated. Treatment with these mycotoxins resulted in decreased cell viability, impaired intestinal barrier function, and altered rates of proliferation and differentiation, including those of enteroendocrine, goblet, and Paneth cell populations. Conclusion This study provides fundamental insights into the growth and differentiation of small intestinal epithelial cells by analyzing timeline-specific organoids using single-cell sequencing. Additionally, it evaluates the toxicity of mycotoxins under conditions that closely resemble those of the small intestine, providing more physiologically relevant data than existing in vitro models and serving as a reliable toxicity assessment model.
Kang et al. (Tue,) studied this question.