Effects of Different Culture Methods on Growth and Keratin 18 Expression of Ruminal Epithelium Cells of Goats

LIU Sile; KANG Jinhe; TAN Zhiliang; WANG Zheng

doi:10.3969/j.issn.1006-267x.2016.04.034

Chinese Journal of Animal Nutrition >

2016 , Vol. 28 >Issue 4: 1225 - 1232

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.04.034

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Effects of Different Culture Methods on Growth and Keratin 18 Expression of Ruminal Epithelium Cells of Goats

LIU Sile ,
KANG Jinhe ,
TAN Zhiliang ,
WANG Zheng

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1. Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Key Laboratory of Agro-Ecological Processes in Subtropical Region of Hunan Province, Changsha 410125, China;
2. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China

Received date: 2015-11-03

Online published: 2016-04-11

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Abstract

This research aimed to study the effects of different culture methods on growth and keratin 18(CK18) expression of ruminal epithelium cells of goats. The test proceeded with ruminal epithelium cells of goats aged 42 days, and the cells were cultured using the methods of enzyme digestion and tissue mass culture. The morphology of cells at the stages of primary culture and subculture was observed under the optical microscope, and the growth curve of the fifth generation of ruminal epithelium cells of goat was tested, meanwhile, ruminal epithelium cells were identified using cell immune fluorescence method. The results showed as follows:1) the primary cultured ruminal epithelium cells of goats obtained by digestion of 0.25% trypsin and 0.02% ethylenediamine tetra-acetate began adherent growth on the 2nd day. There was a significant increase in cell number on the 5th day, and the cell number reached a maximum at the 10th day. 2) The primary cultured ruminal epithelium cells of goats obtained by tissue mass culture began‘escape’from the tissue mass on the 4th day. There was a significant increase in cell number on the 8th day, and the cell number reached a maximum at the 14th day. 3) The CK18 in cytoplasm of cells obtained by the 2 methods exhibited a positive reaction by immune fluorescence staining, and cell purity was higher in tissue mass culture method. 4) Tissue mass culture method expressed significantly higher CK18 compared with enzyme method(P<0.05). In conclusion, ruminal epithelium cells of goats were successfully obtained with high purity using the method of tissue mass culture.

Key words： goat; rumen; epithelium cell; primary culture; isolation; CK18

Cite this article

LIU Sile , KANG Jinhe , TAN Zhiliang , WANG Zheng . Effects of Different Culture Methods on Growth and Keratin 18 Expression of Ruminal Epithelium Cells of Goats[J]. Chinese Journal of Animal Nutrition, 2016 , 28(4) : 1225 -1232 . DOI: 10.3969/j.issn.1006-267x.2016.04.034

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