Blood cell separation method for portunus trituberculatus

A technology of portunus trituratus and a separation method, which is applied to cell dissociation methods, animal cells, invertebrate cells, etc., can solve the problem of not developing crab blood cell separation technology, etc., and achieves simple operation, high separation efficiency, and good activity. Effect
CN108118024AActive Publication Date: 2018-06-05NINGBO UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NINGBO UNIV
Publication Date
2018-06-05

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Abstract

The invention discloses a blood cell separation method for portunus trituberculatus. The method comprises the following steps: diluting iodixanol with an isotonic solution obtained by regulating an osmotic pressure of a PBS buffer solution by utilizing a sodium chloride solution, and performing gradient preparation to obtain three iodixanol solutions of different concentrations; forming gradient solutions of different densities by adopting a cushion technology, and standing to form a continuous density gradient solution to serve as a cell separation solution; collecting blood cells of portunustrituberculatus, and taking mixed cell suspension obtained by diluting the isotonic solution; slowly dropping the mixed cell suspension into the cell separation solution, centrifuging, and dividing the blood cells of portunus trituberculatus into three layers, wherein the uppermost layer refers to clear cells, the intermediate layer refers to small granular cells, and the lowest layer refers to large granular cells. The method has the advantages that only one-step density gradient centrifugation is adopted, the operation is simple, and experiments discover that the separated blood cells haveexcellent viabilities and can be used for cell culture.
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Description

technical field

[0001] The invention relates to a cell separation technology, in particular to a method for separating blood cells of Portunus trituberculatus. Background technique

[0002] Portunus trituberculatus, commonly known as white crab and gun crab, belongs to the subphylum Crustacea, class Malacostraca, order Decapoda, family Portunidae, and the genus Portunus. Widely distributed in sea areas such as China, Japan, North Korea and the Malaysian archipelago, it is an important seafood economic crab in my country. In recent years, Portunus trituberculatus diseases have occurred frequently. According to statistics, among various diseases, pathogenic diseases such as bacteria, viruses and parasites are the main ones, which have caused great losses to my country's aquaculture industry and coastal economy. At present, basic research on the innate immunity of marine crustaceans is gradually increasing to speed up the process of immune prevention and treatment of invertebra...

Claims

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