DF-1 cell culture method used for avian leukemia virus separation

A technology for avian leukosis virus and cell culture, which is applied in the field of DF-1 cell culture for the isolation of avian leukosis virus, can solve the problems of low cell culture efficiency and poor cell culture quality, and achieve the effect of good cell culture quality and rapid cell culture

Inactive Publication Date: 2018-01-05
FOSHAN CITY GAOMING DISTRICT XINGUANG AGRIAND ANIMAL HUSBANDRY
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Problems solved by technology

[0005] However, the existing DF-1 cell culture efficiency is low, and the cell culture quality is poor

Method used

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Embodiment Construction

[0021] Preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0022] The DF-1 cell culture method that a kind of avian leukosis virus isolation of the present invention is used comprises the following steps: 1) the anticoagulant blood 1500rpm centrifugation 10min that collects, absorbs the white villous lymphocyte of supernatant and precipitation junction to 1.5mL In a centrifuge tube, form a plasma sample;

[0023] 2) adding the DF-1 cell suspension to a 96-well cell culture plate, each well has a cell culture solution, the cell culture solution includes 10% FBS and 90% dmem / f-12 medium, and the FBS is fetal bovine serum;

[0024] 3) Separated plasma samples and negative and positive controls were added to each well;

[0025] 4) Put the cell culture plate in the cell cultur...

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Abstract

The invention discloses a DF-1 cell culture method used for avian leukemia virus separation. The method comprises the following steps that 1, centrifugation is conducted on collected anticoagulant blood at 1500 rpm for 10 min, and white villiform lymphocytes at the joint between supernatant and sediments are aspirated into a 1.5 mL centrifuge tube to form a blood plasma sample; 2, a DF-1 cell suspension is added into 96-hole cell culture plate, wherein cell culture liquid exists in each hole; 3, the separated blood plasma sample and positive and negative controls are added into each hole respectively; 4, the cell culture plate is placed into a cell culture box for culture for 24 hours, then the cell culture liquid is replaced by cell maintenance liquid, and culture continues; 5, after 9 days, repeated freezing and thawing is conducted on cells at a temperature of 80 DEG C and room temperature for 2-3 times; 6, after the last freezing and thawing, cultures are collected, and an antigenp27 is detected by means of an ELISA. By means of the method, culture of the DF-1 cells is fast, and the culture quality of the cells is good.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a DF-1 cell culture method for the isolation of avian leukosis virus. Background technique [0002] Avian leukemia is a general term for various neoplastic diseases of poultry caused by viruses of the avian C-type retrovirus group, mainly lymphocytic leukemia, followed by erythroblastic leukemia and myeloblastic leukemia. In addition, it can also cause myeloid cell tumors, connective tissue tumors, epithelial tumors, endothelial tumors, etc. Most tumors invade the hematopoietic system, and a few invade other tissues. [0003] Only chickens can infect the disease under natural conditions. Rous sarcoma virus has the widest host range, and artificial inoculation can also cause tumors in pheasant, guinea fowl, pigeon, quail, turkey and partridge. The resistance of different breeds or strains of chickens to viral infection and tumorigenesis varies greatly. The susceptibility of hens is hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N7/00
Inventor 刘大伟郭露铰冯甫荣张玉龙
Owner FOSHAN CITY GAOMING DISTRICT XINGUANG AGRIAND ANIMAL HUSBANDRY
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