Pass cell line of porcine DC cells as well as establishment method and application thereof
A technology for establishing methods and subculture cells, applied in cell dissociation methods, methods based on microorganisms, animal cells, etc., can solve problems such as inability to pass on for a long time, and achieve the effect of eliminating dependence
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[0081] A method for establishing a passage cell line of pig DC cells, the specific steps are as follows:
[0082] Step S1: Cell Culture
[0083] (1) Separation of mononuclear cells from porcine peripheral blood:
[0084] ① Take fresh EDTA anticoagulated whole blood and dilute the whole blood with an equal volume of diluent or PBS;
[0085] ② Add an appropriate amount of separation liquid to the centrifuge tube. When the diluted blood volume is less than 3 mL, add 3 mL of separation liquid; , otherwise it will affect the separation effect, spread the diluted blood on top of the liquid surface of the separation liquid, and pay attention to keep the interface between the two liquid surfaces clear;
[0086] ③At room temperature, centrifuge with a horizontal rotor of 500-1000g for 20-30min. The larger the volume of blood, the greater the centrifugal force required and the longer the centrifugation time. The best centrifugation conditions depend on the actual situation. The maximu...
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