Abstract: This study aimed to culture in vitro and separate piglet small intestines epithelial cells and observe the soybean agglutinin binding sites. Newborn and health piglets with nonlactation were selected, and the intestinal epithelium cells were cultured and purified. The intestinal epithelium cells purified were identified by immunochemistry method taking the cell keratin 8 antibodies as the first antibody. The intestinal epithelium cells identified were used to mark the soybean agglutinin binding sites with FITCSBA probe by cell agglutinin fluorescent chemistry marker test. The results showed that the porcine small intestinal epithelium cells purified were positive cells by cell keratin 8 antibodies identification, the purity was more than 90%, and the pig small intestine epithelial cells were positively marked by FITCSBA. The porcine small intestinal epithelium cells cultured in vitro posses the soybean agglutinin binding sites, and further specify the porcine small intestinal epithelium cells is one of the main cells combined by soybean agglutinin, which provide the theory for the research on monogastric animal soybean agglutinin antinutrition mechanism.