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Method for establishing mouse model with multiple myeloma combined with chronic graft-versus-host disease

A multiple myeloma and establishment method technology, applied in the field of establishment of multiple myeloma combined with chronic graft-versus-host disease mouse model, to achieve the effects of good repeatability, easy acquisition, and simple process

Active Publication Date: 2021-03-19
GUANGDONG GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most mouse models are cGVHD mouse models or multiple myeloma models, and there are no reports of mouse models involving the combination of the two diseases

Method used

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  • Method for establishing mouse model with multiple myeloma combined with chronic graft-versus-host disease
  • Method for establishing mouse model with multiple myeloma combined with chronic graft-versus-host disease
  • Method for establishing mouse model with multiple myeloma combined with chronic graft-versus-host disease

Examples

Experimental program
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Effect test

Embodiment 1

[0025] This example is an example of establishing the mouse multiple myeloma combined with chronic graft-versus-host disease model of the present invention, and the operation is as follows.

[0026] 1. Mouse preparation

[0027] Specific pathogen Free (SPF) grade NCG (NOD-SCID-IL2rg - / - ) 8-week-old male mice (female mice can also be used) as model mice. The mice in this example were purchased from the Model Animal Research Institute of Nanjing University. The rats were randomly divided into 6 groups, 5 in each group; among them:

[0028] Group A (normal control group): feed synchronously, without intervention;

[0029] Group B (multiple myeloma control group): after whole body irradiation of 100cGy, the suspension of multiple myeloma luciferase reporter gene (Luciferase) stably transfected cell line (IM-9) was injected into NCG mice; Tail vein injection of PRMI1640 culture solution;

[0030] Group C (multiple myeloma+GFP-T control group): After whole-body irradiation of ...

Embodiment 2

[0052] This example is an example of in vivo imaging of tumor burden in mice with multiple myeloma combined with cGVHD.

[0053] 1. For IM-9 cells, due to the expression of luciferase, imaging can be performed through the substrate to detect the residual and growth status of IM-9 cells;

[0054] 2. Inject the substrate D-luciferin 200ul (15mg / ml) through the intraperitoneal cavity, and after 5 minutes of timing, put it into the anesthesia room attached to the in vivo imager, and perform anesthesia with isoflurane at the same time;

[0055] 3. Put the anesthetized mouse into the imager for imaging, and perform imaging in the automatic exposure mode (that is, the exposure time will be automatically adjusted according to the signal strength);

[0056] 4. After imaging, put the mice back into the cage for feeding;

[0057] 5. Data analysis.

[0058] like image 3 Shown: A is the in vivo imaging of mice on the 7th day after the tumor was planted, and all the tumors in the mice w...

Embodiment 3

[0060] This example is an example of flow cytometry detection of tumor burden in mice with multiple myeloma complicated with cGVHD.

[0061] 1. For the IM-9 cell line, because it expresses the CS1 antigen, the expression of the CS1 antigen in the peripheral blood can be detected by flow cytometry, and the residual situation of the IM-9 cells can be detected;

[0062] 2. Collect blood from tail vein of mice in multiple myeloma+CS1CAR-T group;

[0063] 3. Stain the peripheral blood of the mouse with anti-human CS1 for 30 minutes. After 30 minutes, add 500ul of erythrocyte lysate into the flow tube, lyse for 10 minutes, add 3ml of PBS to wash, centrifuge at 300g for 5min, wash with PBS again, add 200ul PBS spare.

[0064] 4. Detect the expression of CS1 antigen on the computer, and analyze the data.

[0065] like Figure 4 Shown: the expression of CS1 antibody in the mice of group D, CS1 in the figure further proves the tumor burden of the mice, and the residual multiple myelo...

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Abstract

The invention provides a method for establishing a mouse model with multiple myeloma combined with chronic graft-versus-host disease (cGVHD). The method sequentially comprises the following steps of taking an NCG (NOD-SCID-IL2rg- / -) mouse as a model mouse; after the mouse receives 100 cGy of whole-body irradiation, injecting a suspension of multiple myeloma luciferase reporter gene cell strain (IM-9) into the mouse; and 8 days after the mouse is injected, injecting a CS1CAR-T cell suspension into the body of the mouse through a caudal vein. The modeling method of the model has the characteristic of simulating the occurrence and development of the chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation of a patient suffering from multiple myeloma clinically; and the method is simple and feasible in process, high in modeling efficiency and good in repeatability, typical pathological changes occur in the mouse, and an experimental platform is provided for research on prevention and treatment of human multiple myeloma combined with the cGVHD after transplantation in living animals.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for establishing a mouse model of multiple myeloma combined with chronic graft-versus-host disease. Background technique [0002] Chronic graft-versus-host disease is the main complication of long-term survival patients after allogeneic hematopoietic stem cell transplantation. It not only seriously affects the quality of life of patients, but also is the main cause of death. It is of great practical significance to further clarify the mechanism of cGVHD, explore the relationship between GVL and GVHD, and study active and effective treatment methods to reduce or alleviate its occurrence and disease course. Therefore, the establishment of combined multiple myeloma and cGVHD animal models is an important basis for translational medical research. At present, most mouse models are cGVHD mouse models or multiple myeloma models, and there is no report of a mouse model involving the ...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N15/867C12N5/10C12N5/09
CPCA01K67/0271C12N15/86C12N5/0693A01K2227/105A01K2267/0331C12N2740/10043C12N2510/00
Inventor 翁建宇陈晓梅杜欣赖沛龙王玉连耿素霞黄欣曾令基
Owner GUANGDONG GENERAL HOSPITAL
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