几种酶分离山羊小肠上皮细胞方法的比较

Comparison of Methods to Generate Vialde Goat Intestinal Epithelial Call by Enzymes

用不同酶消化分离山羊小肠上皮细胞,从而找到最佳的分离方法,为进一步研究IEC提供合适的细胞模型。本研究分别用胶原酶Ⅰ、嗜热菌蛋白酶、胶原酶Ⅺ/中性蛋白酶Ⅰ消化分离刚出生未进奶的山羊小肠上皮细胞,通过观察IEC生长状态、形态特征、抗原鉴定比较哪种分离效果最佳。结果表明,联合用胶原酶Ⅺ/中性蛋白酶Ⅰ1h可分离出大量健全绒毛隐窝单位,进一步培养获得的小肠粘膜上皮(IEC),上皮细胞膜抗原均呈阳性,用胶原酶Ⅰ和嗜热菌蛋白酶分离得到的大部分是成纤维细胞。选用用胶原酶Ⅺ/中性蛋白酶Ⅰ作为分离IEC的消化酶,可获得较纯的IEC。

To find the optimal method to isolate and culture the goat intestine epithelial cells,collagenase I . thermolysin.collagenaseXI/dispase I digestion technique were used to isolate IEC from the fetal goat. Optimal culture conditions were determined by both cellular morphology under light microscope and 4 immunocytochemical characterization. The results showed that large amount of villi crypt units were attainable and the attached cells were positive for cytokeratin using collagenaseXI/dispase I digestion technique while many fibroblast cells contaminated when using collagenase I and thermolysin digestion techniques. This study showed that viable epithelial cells can be relatively easily generated from fetal goat small intestine by collagenaseXI/dispase I digestion technique.