Characteristics of Ruminal Epithelium Cell Cycle Distribution, Proliferation and Apoptosis of Liuyang Black Goats

HAN Qipeng; LUO Ling; JIE Hongdong; WANG Kaijun; ZHANG Peihua; ZHOU Chuanshe; KONG Zhiwei; TANG Shaohui

doi:10.3969/j.issn.1006-267x.2016.07.034

Chinese Journal of Animal Nutrition >

2016 , Vol. 28 >Issue 7: 2269 - 2277

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.07.034

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Characteristics of Ruminal Epithelium Cell Cycle Distribution, Proliferation and Apoptosis of Liuyang Black Goats

HAN Qipeng ,
LUO Ling ,
JIE Hongdong ,
WANG Kaijun ,
ZHANG Peihua ,
ZHOU Chuanshe ,
KONG Zhiwei ,
TANG Shaohui

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1. Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;
2. Key Laboratory for Agro-Ecological Processes in Subtropical Region, Hunan Research Center of Livestock & Poultry Sciences, South-Central Experimental Station of Animal Nutrition and Feed Science of Ministry of Agriculture, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China

Received date: 2016-01-25

Online published: 2016-07-28

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Abstract

The present study was carried out to establish an in vitro culture model of rumen epithelial cells of Liuyang black goats and investigated cell cycle distribution, proliferation and apoptosis. Ruminal epithelium tissue of 60-day-old Liuyang black goats was collected, and digested with 0.25% trypsin and 0.02% ethylene diamine tetraacetic acid (EDTA) to obtain individual primary cultured ruminal epithelium cells for in vitro culture. The morphous of ruminal epithelial cells in primary culture and subculture was observed under inverted microscope, the growth activity curve of cells was tested by cell number count assay, the identification of subcultured ruminal epithelium cells was carried out using the method of immunohistochemistry, and the subcultured ruminal epithelium cells cycle distribution and apoptosis ratio were determined through flow cytometry method. The results showed as follows: 1) the primary cultured ruminal epithelium cells of goats were obtained by digestion of 0.25% trypsin and 0.02% EDTA, began adherent growth after cultured for 1 d, an exponential growth started and resembled a "hill" in appearance since 2 d, the cell growth rate reached the highest at 3 to 4 d, and got stable at 7 d. 2) Identified by immunohistochemistry method, the cytoplasm was brown, which meant that cytokeratin 19 (CK19) was positive expressed. 3) The combination staining of Annexin V and propidium iodide showed that the apoptosis ratio of cells significantly increased with the prolongation of culture time (P<0.01). In conclusion, the ruminal epithelium cells of Liuyang black goats are obtained successfully by the method of 0.25% trypsin+0.02% EDTA digestion, which provided cell model for further study on mechanism and the function of ruminant rumen.

Key words： Liuyang black goat; ruminal epithelium cell; cycle distribution; apoptosis

Cite this article

HAN Qipeng , LUO Ling , JIE Hongdong , WANG Kaijun , ZHANG Peihua , ZHOU Chuanshe , KONG Zhiwei , TANG Shaohui . Characteristics of Ruminal Epithelium Cell Cycle Distribution, Proliferation and Apoptosis of Liuyang Black Goats[J]. Chinese Journal of Animal Nutrition, 2016 , 28(7) : 2269 -2277 . DOI: 10.3969/j.issn.1006-267x.2016.07.034

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