体外培养猪小肠上皮细胞大豆凝集素结合位点的标记

In vitro Marker of Soybean Agglutinin Binding Sites in Porcine Intestinal Epithelial Cells

本研究旨在获得体外培养状态下的猪小肠上皮细胞,研究猪小肠上皮细胞与大豆凝集素的结合情况。选取新生未哺乳健康仔猪,取小肠组织对小肠上皮细胞进行分离培养,并进行纯化。纯化后的细胞以角蛋白抗体-8为一抗进行细胞免疫化学试验鉴定,鉴定后的猪小肠上皮细胞以FITC-SBA为探针进行细胞凝集素荧光标记化学试验,对猪小肠上皮细胞大豆凝集素结合位点进行标记。结果表明:试验分离纯化的细胞经角蛋白抗体-8鉴定为阳性,所分离纯化的细胞为猪小肠上皮细胞,经检测其纯度达90%以上,猪小肠上皮细胞FITC-SBA标记结果为阳性。体外培养的猪小肠上皮细胞表面存在大量的大豆凝集素结合位点,进一步明确了猪小肠上皮细胞是大豆凝集素结合的主要细胞类型之一,为单胃动物大豆凝集素抗营养机制研究提供理论依据。

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.