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Separation method of bovine blood CD4 + T lymphocytes

A lymphocyte and separation method technology, which is applied in the direction of cell dissociation method, blood/immune system cells, animal cells, etc., can solve the problem of no report of T lymphocytes, etc., and achieve the effect of short separation time, low damage and high purity

Pending Publication Date: 2022-03-18
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] At present, the research on bovine acquired immune function is limited to the report on the lymphocyte level, but the bovine blood CD4 + T lymphocytes have not been reported

Method used

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  • Separation method of bovine blood CD4 + T lymphocytes

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Embodiment Construction

[0020] A method for isolating bovine blood CD4+ T lymphocytes, comprising the following steps:

[0021] Step (1), take 15-20 mL of fresh anticoagulated whole blood from the oxtail vein (conventional anticoagulated blood collection tubes on the market can be used), and use an equal volume of whole blood and tissue diluent (Solabo bovine peripheral blood lymphocyte separation diluent contained in the kit) to dilute the whole blood to obtain a whole blood dilution;

[0022] Step (2), add 5 mL of separation liquid (separation liquid contained in Solebo Bovine Peripheral Blood Lymphocyte Separation Kit) and 5 mL of whole blood dilution obtained in step (1) to a 15 mL centrifuge tube, and dilute the Spread the whole blood dilution described above on the surface of the separation liquid (note that the interface between the two liquid surfaces is clear); at room temperature, centrifuge at 950g in a horizontal rotor for 30 min, and absorb the buffy coat layer to obtain lymphocytes cont...

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Abstract

The invention discloses a separation method of bovine blood CD4 + T lymphocytes, and relates to the field of bioengineering.The separation method comprises the following steps that oxtail vein blood is collected; separating lymphocytes by using a commercial lymphocyte separation kit; centrifuging, collecting precipitate, and adding a red blood cell lysis solution to lyse red blood cells; adding a culture medium, washing, centrifuging and collecting precipitate; after cell counting, adding a primary antibody of CD4 + T lymphocytes for incubation; adding a magnetic bead secondary antibody for incubation; and finally, separating the CD4 + T lymphocytes by using a magnetic column. The invention introduces the bovine CD4 + T lymphocyte separation method for the first time, and fills the blank of the test method technology in the field. The bovine CD4 < + > T lymphocytes obtained by the method are good in activity, large in quantity and high in purity. The kit can be widely applied to clinical diagnosis besides scientific research, and has a good market prospect.

Description

Technical field: [0001] The invention relates to the field of bioengineering, in particular to a bovine blood CD4 + Methods of isolation of T lymphocytes. Background technique: [0002] As a kind of effector cells, lymphocytes widely exist in peripheral blood, bone marrow, umbilical cord blood or immune organs of animals. The acquisition of high-purity lymphocytes is of great significance for the cytological diagnosis of diseases, component blood transfusion, and the preparation of therapeutic cell products. T cells are a subset of lymphocytes, including CD8 + and CD4 + (T helper, Th) T cells. Further divisions of Th cells include Th1, Th2, Th17 and T regulatory cells (Treg). The balance between pro-inflammatory Th17 cells and anti-inflammatory Treg cells determines the extent of the immune response, which is critical for homeostasis and health in an organism. In clinical testing, the examination of specific cell functions and biological characteristics often involves...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2509/00C12N2501/505
Inventor 徐闯李铭张冰冰杨威尹钰峰何雨欣温佳楠王晶晶
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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