Method for extracting plastocyte of mouse

An extraction method and platelet technology, applied in biochemical equipment and methods, blood/immune system cells, sedimentation by centrifugal force, etc., can solve problems such as poor purity and insufficient platelet concentration

Inactive Publication Date: 2009-04-08
LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0003] In view of the shortcomings of the traditional platelet extraction method, this patent provides a mouse platelet extraction method to solve the problems of insufficient concentration and poor purity of the extracted platelets

Method used

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Examples

Experimental program
Comparison scheme
Effect test

experiment Embodiment 1

[0009] From October 10th to October 25th, 2007, 80 SPF grade BALB / c mice were used to extract platelets using this method. The purity of platelets reached 97%, and the concentration was 1×10 10 , after using the platelets to prepare immune serum, a model of allergic purpura was prepared, and 95% of the models were formed, and the model was used to evaluate the curative effect and mechanism of anti-allergic purpura traditional Chinese medicine.

experiment Embodiment 2

[0011] From April 15th to April 30th, 2008, 100 SPF grade BALB / c mice were used to extract platelets using this method. The purity of platelets reached 95%, and the concentration was 3×10 10 , after using the platelets to prepare immune serum, a model of allergic purpura was prepared, and 96% of the models were formed, and the model was used to evaluate the curative effect and mechanism of anti-allergic purpura traditional Chinese medicine.

experiment Embodiment 3

[0013] From June 12th to June 27th, 2008, 120 SPF grade BALB / c mice were used to extract platelets using this method. The purity of platelets reached 98%, and the concentration was 5×10 10 , after using the platelets to prepare immune serum, prepare anaphylactoid purpura model, 98% of the model is formed, and use this model to evaluate the curative effect and mechanism of anti-allergic purpura traditional Chinese medicine.

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Abstract

The invention relates to an extraction method of mouse platelet. Blood drawn from orbita vein of mouse is placed into an anticoagulation centrifugal tube, centrifugated at the speed of 800rmp for 10min after standing for 30min, and the extracted supernatant fluid is plasma rich in platelet. The supernatant fluid is placed in a clean test tube and centrifugated at the speed of 3500rpm for 10min. After filtering the supernatant fluid, the sediments at the bottom of the tube are the platelets. 50ul to 100ul of ammonium oxalate solution with a mass concentration of 1 percent is added into the test tube and slightly stirred with a glass rod, and then 2ml to 3ml of ammonium oxalate solution with a mass concentration of 1 percent is added, and stood for 5min, to make sure the hematid is dissolved; the hematid is centrifugated at the speed of 3500rpm for 10min, after the supernatant fluid is abandoned, the test tube is added with a few platelet scrub solution and blown and beaten to turn the platelet into a platelet suspension. The suspension is centrifugated at the speed of 3300rpm for 10min, removed the supernatant in the suspension, the test tube is added with a few platelet scrub solution and blown and beaten to turn the platelet into a platelet suspension. The platelet is used as an antigen to immunify guinea pigs, and the serum of such guines pigs is injected into the mouse to prepare allergic purpura. The preparation method has simple operation, can obtain high concentration platelet, and can comparatively better solve the problem of impure platelet.

Description

Technical field: [0001] The invention relates to a method for extracting mouse platelets. The extracted platelets are used as antigens to immunize guinea pigs, and the immune serum of the guinea pigs will be used to prepare mouse idiopathic thrombocytopenic purpura animal models. Background technique: [0002] The pathogenesis of idiopathic thrombocytopenic purpura (ITP) is closely related to immunization, and the ITP model induced by immunization is highly similar to human ITP in terms of pathological characteristics and healing mechanism, so the experimental research mainly uses immunization to establish animal models, small The mouse ITP model is highly sensitive to various anti-ITP drugs, so the successful preparation of the mouse ITP model is one of the key technologies for the research of anti-ITP drugs. However, a significant problem with this modeling method is that the purity of the antigen is not enough, that is, the purity of the platelets isolated from the whole ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A61B19/00B01D21/26C12N5/078
Inventor 林庶茹才丽平范颖曲道炜李阳曲怡蒋宁
Owner LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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