Application of MDSCs in constructing BA animal model and construction method of BA mouse model

A mouse model and construction method technology, which can be used in medical preparations containing active ingredients, pharmaceutical formulations, unknown raw materials, etc., can solve the problems of low RRV detection rate and lack of convincing.

Active Publication Date: 2019-04-26
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the most obvious flaw is that the model is induced by RRV, but the detection rate of RRV in clinical BA liver

Method used

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  • Application of MDSCs in constructing BA animal model and construction method of BA mouse model
  • Application of MDSCs in constructing BA animal model and construction method of BA mouse model
  • Application of MDSCs in constructing BA animal model and construction method of BA mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Reagent consumables

[0064] RPMI 1640 medium (11875093, Gibco, USA), red blood cell lysate (RT122, TIANGEN, CHN), trypan blue (T8154, Sigma, USA), mouse MDSC magnetic bead sorting kit (130-094-538, Miltebyi, USA), 40 μm cell sieve (258369, NEST), 15 ml centrifuge tube (352097, CORNING), 1 mL insulin syringe (9161635S, BD), cell counting plate (145-0011, BIORAD).

Embodiment 2

[0065] Embodiment 2 constructs the BA model induced by RRV

[0066] Balb / c newborn mice, within 24 hours after birth, each newborn mouse was intraperitoneally injected with rhesus monkey rotavirus (MMU18006 virus strain, the titer of the virus suspension was 1×10 6 PFU), the injection dose is 20 μL.

Embodiment 3

[0067] Embodiment 3 sorting MDSC (Gr-1 + CD11b + )cell

[0068] (1) Steps for separating the liver tissue suspension:

[0069] Put the 90mm culture medium on crushed ice and pour about 10mL of 1640 medium into it; kill the mouse in Example 2 by cervical dislocation, soak and disinfect with 75% alcohol, fix the mouse on the foam board, and use tweezers and scissors The liver was bluntly separated, put into a 40 μm cell sieve, and gently ground with a syringe with glue until there were no large tissue pieces; the ground cell suspension was then passed through a 40 μm cell sieve to filter out large tissue pieces, and the sieved Transfer the cell suspension to a 15mL centrifuge tube, centrifuge at 4°C 1000rpm for 3 minutes; discard the supernatant, add 2mL red blood cell lysate, react on ice for 1 minute, add 8mL1640 medium to terminate, and centrifuge again at 4°C 1000rpm for 3 minutes; discard Remove the supernatant, add 2mL 1640 medium to resuspend the cells, gently pipette,...

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Abstract

The invention relates to the technical field of animal model construction, in particular to an application of MDSCs in constructing a BA animal model and a construction method of a BA mouse model. Theinventor finds through a test that MDSCs plays an important role in BA and can be used for constructing the BA animal model. The construction method of the BA mouse model adopts a neonatal mouse by using MDSCs and adopts non-viral infection, so that the method is simple and safe and the modeling success rate is close to 100%, the mouse xanthochromia appears on the fourth day of birth and jaundiceappears early. The liver pathological section result shows that the symptom is as same as that of BA and no RRV is left in liver. The model can be used for nosetiologic research of BA and research ofdiagnosis and treatment methods.

Description

technical field [0001] The present invention relates to the technical field of animal model construction, in particular to an application of MDSCs in the construction of BA animal models and a construction method of BA mouse models. Background technique [0002] Biliary atresia (BA) is a common disease causing obstructive jaundice in newborns, with poor prognosis and high mortality. The etiology is still unknown. The basic pathological changes are progressive inflammation of extrahepatic and intrahepatic bile ducts and liver fibrosis. my country is an area with a high incidence of BA disease, with an incidence rate of 1 / 5000-1 / 10000. Most of the children were found 1-3 months after birth, and most of them died of liver failure within 1.5 years of age because they could not perform liver transplantation. At present, due to the lack of suitable animal models, no breakthroughs have been seen in the etiology research, diagnosis and treatment of the disease. Therefore, it is p...

Claims

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Application Information

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IPC IPC(8): A01K67/02A61K35/407
CPCA01K67/02A61K35/407
Inventor 林泽锋张锐忠陈严付铭王哲林辉婷童燕陆钟微余家康夏慧敏
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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