Isolation, Purification, Culture and Identification of Goat Pancreatic Acinar Cells in Vitro

ZHANG Tianxi; YAN Aifei; YANG Chao; CHENG Yan; HE Zhixiong; TAN Zhiliang

doi:10.3969/j.issn.1006-267x.2022.09.061

Chinese Journal of Animal Nutrition >

2022 , Vol. 34 >Issue 9: 6088 - 6096

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.09.061

RESEARCH PAPER

Isolation, Purification, Culture and Identification of Goat Pancreatic Acinar Cells in Vitro

ZHANG Tianxi ,
YAN Aifei ,
YANG Chao ,
CHENG Yan ,
HE Zhixiong ,
TAN Zhiliang

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1. Public Technology Center of Institute of Subtropical Agriculture, Chinese Academy of Sciences, Hunan Research Center of Livestock & Poultry Sciences, National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Key Laboratory for Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received date: 2022-03-22

Online published: 2022-10-17

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Abstract

The purpose of this study was to establish a primary culture method of pancreatic acinar cells in Xiangdong black goats, and to provide a cell model for the study of pancreatic exocrine function in ruminants. In this experiment, collagenase Ⅳ was used to isolate goat pancreatic acinar cells, and the optimum concentration and digestion time of collagenase Ⅳ, comparison with 0.25% trypsin digestion efficiency, purification and identification of acinar cells were studied. The results showed as follows:1) when 1.0 mg/mL collagenase Ⅳ was used to digest 20 min in the pancreatic tissue of Xiangdong black goats, the isolation effect of acinar cells was the best. The total cell number of 2.275×10⁶ cells/mL and the cell viability of 91% were obtained by digestion under this condition. 2) The primary cells were purified by differential digestion, differential adhesion and cell scraping, and the goat pancreatic acinar cells with good activity and "cobblestone" shape were obtained. 3) After purification, the acinar cells can be stably passed on for more than 5 generations, and the morphology does not change. 4) The rabbit anti-carboxypeptidase A1 (CPA1) was used as the first antibody to identify the first and fifth generation goat pancreatic acinar cells by immunofluorescence technique, and the results were positive. To sum up, the goat pancreatic acinar cells are quickly isolated by this method, and the isolated goat pancreatic acinar cells are successfully purified, identified and stably cultured.

Key words： goats; pancreatic exocrine function; pancreatic acinar cells; cell culture

Cite this article

ZHANG Tianxi , YAN Aifei , YANG Chao , CHENG Yan , HE Zhixiong , TAN Zhiliang . Isolation, Purification, Culture and Identification of Goat Pancreatic Acinar Cells in Vitro[J]. Chinese Journal of Animal Nutrition, 2022 , 34(9) : 6088 -6096 . DOI: 10.3969/j.issn.1006-267x.2022.09.061

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