Isolation Culture and Identification of Porcine Intestinal Epithelial Cells

ZHAN Kang, ZUO Xiaoxin, CHEN Yinyin, GONG Xiaoxiao, ZHAN Jinshun, ZHAO Guoqi

doi:10.3969/j.issn.1006-267x.2015.05.018

Chinese Journal of Animal Nutrition >

2015 , Vol. 27 >Issue 5: 1477 - 1484

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.05.018

Experimental Methods

Isolation Culture and Identification of Porcine Intestinal Epithelial Cells

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College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

Received date: 2014-12-15

Online published: 2015-05-13

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Abstract

This experiment was designed to establish a functional immortalized porcine intestinal epithelial cells line and study porcine intestinal nutrient absorption, immune regulation and the pathogenesis of the neonatal porcine intestinal disease, which provided the cell model. Tissue culture method was used in this experiment to separate and purify the porcine epithelial cells. And the methods of cytokeratine 18 immunofluorescence, cell proliferation curve and karyotype analysis were used to identify the porcine intestinal epithelial cells. The results showed that the porcine intestinal epithelial cells could be successfully cultured and passed 11 generations stably by tissue culture method. 2) The porcine intestinal epithelial cells showed positive reaction against cytokeratine 18 antigen, and the karyotype was diploid. 3) The 11th generation cells still have characteristics of epithelial cells, showing "cobblestone-shape" and epithelial-like morphological features. However, the porcine intestinal epithelial cells were out of shape and went to the procedure of apoptosis after 11 generations. Most of them went to death in 15th generation and seldom were adherent to the dishes anymore. In summary, the experiment succeed to obtain a porcine intestinal epithelial cells line with stable biological functions by tissue culture method, and they pass 11 generations stably, which provided the experimental materials for immortalized cells.

Key words： porcine intestinal epithelial cell; function; intestinal nutrition and immunity; immortalize

Cite this article

ZHAN Kang, ZUO Xiaoxin, CHEN Yinyin, GONG Xiaoxiao, ZHAN Jinshun, ZHAO Guoqi . Isolation Culture and Identification of Porcine Intestinal Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2015 , 27(5) : 1477 -1484 . DOI: 10.3969/j.issn.1006-267x.2015.05.018

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