<p>Drug absorption through the intestinal epithelium is a major process that orally administered drugs must pass through. Current well-based in vitro cell-based models do not accurately simulate the drug absorption process, and chip-based absorption models have been gaining attention as an alternative to the conventional drug absorption models. We developed a gut-on-a-chip to observe drug absorption by culturing enterocytes on a previously developed multi-organ-on-a-chip (MOC). First, to demonstrate the feasibility of culturing enterocytes on a chip, we selected and cultured two types of enterocytes and observed their survival rate and morphology. It was confirmed that the physiological functions of the enterocytes were appropriately expressed by observing the expression of intestine-related proteins and enzyme activities of the two enterocytes cultured on the chip. A comparison of the static well-based environment and the dynamic chip environment using two types of enterocytes showed that the overall intestinal function was improved in the chip environment compared to the well environment, and human intestinal epithelial primary cells (HIEP) were physiologically superior to intestinal epithelial cell line (Caco-2). We examined drug absorption properties to confirm the possibility of gut-on-a-chip being used as a drug absorption model. We conclude that gut-on-a-chip containing HIEP cells is superior to the conventional well-based gut model containing Caco-2 cells. We hope that the gut-on-a-chip developed in this study can potentially be used as a new and more accurate intestinal model and intestinal absorption model.</p>