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Separating and culturing method of primary duck liver cells

A culture method, duck liver technology, which is applied in the field of cell culture of modern biotechnology, can solve the problems of poor activity and small number of liver cells, and achieve the effect of stable quality, high purity and large number of cells

Inactive Publication Date: 2018-11-02
JIANGYIN CHI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mechanical method is simple to operate, but the number of isolated hepatocytes is small and the activity is poor

Method used

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  • Separating and culturing method of primary duck liver cells
  • Separating and culturing method of primary duck liver cells

Examples

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Embodiment Construction

[0023] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0024] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0025] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0026] The experimental instruments and reagents used in this experiment are as follows:

[0027] A set of surgical instruments, CCK-8 cell counting box (Sigma, USA), inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifuge (TD24B-WS, Shanghai), ultra-low temperature refrigerator ( Zhongke Meiling), pipette gun (Eppendorf, USA), electronic analytical balance (Sartorius, USA), ultra-clean bench (HJ-CJ-1D, Shanghai), CO 2 Cell incub...

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Abstract

The invention provides a separating and culturing method of primary duck liver cells. The method comprises the following steps: (1) selecting male Chongqing sheldrake at the age of 1-2 months, and injecting pentobarbital sodium and heparin into enterocoelia; (2) fixing the narcotized sheldrake to a board with venter upward, and excising the enterocoelia in a sterile condition; (3) performing venous cannula, and pouring reheated HBSS buffering liquid through a constant-flow pump; (4) pouring preheated GBSS buffering liquid through the constant-flow pump; (5) pouring the preheated GBSS mixed enzyme solution through the constant-flow pump; (6) shearing the GBSS mixed enzyme solution for tissue; oscillating and digesting; filtering through a sieving net; centrifuging; and removing the supernatant; (7) detecting the cell activity of the cell filtrate through trypan blue staining; (8) counting the cells, and suspending and inoculating to a cladded culture flask through a complete culture medium; and culturing in an environment that the temperature of 37 DEG C, and 5% of CO2 is added; (9) identifying the cell forms; (10) performing through ELISA; and (11) performing RT-PCR detection. Theseparating and culturing method of the duck liver cells is simple to operate; a large amount of cells can be obtained; the survival rate is high; the method is an ideal separating and culturing methodof the primary duck liver cells; and reliable cell resources are provided to test.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and in particular relates to a method for separating and culturing primary duck liver cells. Background technique [0002] Duck hepatitis B virus (DHBV) and human hepatitis B virus (HBV) belong to the family Hepadnaviridae. Due to the similarity between DHBV and HBV, the DHBV-infected duck model has been widely used in animal experimental models in the study of HBV pathogenic mechanism and its treatment. DHBV infection can be used as an experimental animal model for the pathogenesis of liver disease caused by HBV infection and for screening and treatment of human viral hepatitis drugs. Studying DHBV infection can indirectly recognize HBV. The culture method of primary duck liver cells established in the present invention is an important tool for studying HBV, and provides good in vitro culture conditions for screening antiviral drugs. [0003] At present, the separa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2501/33
Inventor 不公告发明人
Owner JIANGYIN CHI SCI
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