Rapid separating method for cyprinid fish hepatocytes

Abstract

The invention discloses a rapid separating method for cyprinid fish hepatocytes. The rapid separating method comprises the steps of chopping fish livers to obtain fish liver fragments and adding a Hank's balanced salt solution into the fish liver fragments, and performing oscillation and centrifugation to obtain precipitates; adding digestive juice consisting of CaCl2, bovine serum albumin, collagenase, dispase, a trypsin inhibitor, deoxyribonuclease I and the Hank's balanced salt solution into the precipitates, and performing oscillation, beating and sieving with a cell strainer to obtain large tissue precipitates and a cell suspension; adding a cleaning solution consisting of the CaCl2, the bovine serum albumin and the Hank's balanced salt solution into the large tissue precipitates, andperforming beating and sieving with the cell strainer to obtain a cell suspension, repeating the step for 3 times to obtain a merged cell suspension, performing centrifugation on the merged cell suspension to obtain cell precipitates, adding the cleaning solution into the cell precipitates, and performing beating and centrifugation to prepare primary hepatocytes. The method effectively solves theproblems that cost is high and the sizes of fishes are required to be large due to the fact that the separating time is long and multiple enzyme preparations are required by employing an existing method

Description

technical field [0001] The invention relates to the technical field of fish liver cell separation, in particular to a rapid separation method for carp fish liver cells. Background technique [0002] The liver is one of the most important organs in the body. It has the functions of digestion, metabolism and immune regulation, and is the target organ of harmful environmental factors. Therefore, primary cells isolated from the liver are often used as the object of research in pharmacology, toxicology, immunology, cell biology and endocrinology. Fish hepatocytes have also been isolated and cultured, and they have been used in the study of fish liver structure and function, such as temperature acclimation of fish, cell senescence, cell interaction, lipoprotein metabolism, nutrient metabolism and endocrine regulation, vitellogenin Synthesis, hormone receptor expression, metabolism and toxicology studies of cytochrome P450. [0003] The perfusion method is often used to isolate h...

AI Technical Summary

Problems solved by technology

[0004] Aiming at the above-mentioned deficiencies in the prior art, the present invention provides a rapid separation and culture method of carps hepatocytes, which can effectively solve the problems of long separation time and many enzyme preparations required in the existing methods. Lead to high cost and large fish volume requirements