A method for the isolation and primary culture of grass carp dendritic cells

A dendritic cell and primary culture technology, which is applied in the field of grass carp dendritic cell isolation and primary culture, can solve problems such as the method of isolating and culturing grass carp dendritic cells that have not yet been seen, and reduce the risk of cell contamination Probability, less damage to the donor, and easy access to the effect
CN109294989BActive Publication Date: 2021-11-09HUAZHONG AGRI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
HUAZHONG AGRI UNIV
Publication Date
2021-11-09

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Abstract

The present invention provides a method for the isolation and primary culture of grass carp dendritic cells, comprising the following steps: Step 1, the separation of grass carp spleen and head kidney: the grass carp is anesthetized to death, the spleen and head kidney are taken out, and the attached connective tissue is removed and adipose tissue and soaked; step 2, preparation of grass carp spleen and head kidney single cell suspension: continuous sieve filtration to obtain cell suspension; step 3, separation of grass carp spleen and head kidney mononuclear cells: Ficoll separation, after centrifugation Absorb and wash the cells of the buffy coat; step 4, static culture: adjust the concentration of monocytes, and place them in an incubator for culture; step 5, enrichment and purification of grass carp dendritic cells: collect suspension cells after culturing for a period of time, and use Grass carp dendritic cells were purified by density gradient centrifugation. The method can effectively prepare a large amount of grass carp dendritic cells, and the purity of the obtained grass carp dendritic cells can reach more than 80 percent; the operation process is simple and clear, and the time consumption is short.
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Description

technical field

[0001] The invention relates to the technical field of in vitro culture of fish cells, in particular to a method for the isolation and primary culture of grass carp dendritic cells. Background technique

[0002] As an important technology of cell biology and even biological research, cell culture occupies an important position in the field of biological research. Animal tissue (cell) culture began in the early 20th century, and has become a widely used technique in biological and medical research and applications. Fish cell culture started relatively late, since the 1960s. Mainly used in virology, immunology and physiological and toxicology research. The isolation and culture of fish dendritic cells is of great significance to the prevention, diagnosis and treatment of fish diseases.

[0003] Dendritic cell (DC) is the most powerful antigen presenting cell (Antigen presenting cell, APC) known in vivo (Steinman, 1991). DC is a key factor in identifying inte...

Claims

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