Method for constructing humanized Ph chromosome positive acute lymphocytic leukemia mouse model

An acute lymphocyte and mouse model technology, applied in the biological field, can solve the problems of high cost, not allowing breeding, cumbersome mouse procedures, etc., and achieve the effect of high efficiency, good repeatability, and simple and easy modeling process

Inactive Publication Date: 2014-04-23
PEOPLES HOSPITAL PEKING UNIV
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Problems solved by technology

[0004] In 2004, Dr. Shultz of the Jackson Laboratory in the United States took the lead in establishing the most advanced NOD/SCID/IL2Rγ in the world null Immunodeficiency mouse model, due to IL-2Rγ deficiency in this model, the signal transduction pathway that binds to common cytokine receptors with high affinity is inhibited, resulting in various natural immune function defects such as NK cells
[0005] However, domestic NOD/SCID/IL2rγ null The source of mice can only be purchased from the United States or Japan at present. Not only is the cost high, but the procedures for mouse purchase, customs declaration, and isolation and quarantine are cumbersome, and only single-sex NOD/SCID/IL2rγ can be purchased null Mice are not allowed to breed in China, which severely restricts NOD/SCID/IL2rγ

Method used

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  • Method for constructing humanized Ph chromosome positive acute lymphocytic leukemia mouse model
  • Method for constructing humanized Ph chromosome positive acute lymphocytic leukemia mouse model
  • Method for constructing humanized Ph chromosome positive acute lymphocytic leukemia mouse model

Examples

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Embodiment 1

[0029] Example 1 Construction of humanized Ph + ALL mouse model

[0030] In this embodiment, a humanized Ph chromosome-positive acute lymphoblastic leukemia mouse model is constructed with a preferred dose, including the following steps:

[0031] Step A, Ph + Preparation of bone marrow mononuclear cells from patients with ALL: Ph + Bone marrow samples from patients with ALL were anticoagulated with heparin, and bone marrow mononuclear cells were separated and prepared using human lymphocyte separation medium, and RPMI1640 culture medium containing 10% fetal bovine serum was used Resuspended cells to a final cell density of 5 x 10 8 / mL, Ph + Cell suspension of ALL bone marrow mononuclear cells. Specific steps are as follows:

[0032] 1. Add 5mL lymphocyte separation medium to a 15mL centrifuge tube;

[0033] 2. Take 10 mL of heparin-anticoagulated human bone marrow and mix it well with an equal amount of Hank's solution, slowly add it to the layered liquid surface...

Embodiment 2

[0046] Different from Example 1, the final cell density in step A was 8×10 8 / mL;

[0047] Step B, taking NOD / SCID female mice that were 6 weeks old one week before pretreatment, 60 Co irradiation dose was 2.4 cGy, and 250 μg of anti-mouse CD122 monoclonal antibody was injected per mouse.

Embodiment 3

[0049] Different from Example 1, the final cell density in step A was 1×10 8 / mL;

[0050] Step B, taking NOD / SCID female mice that were 6 weeks old one week before pretreatment, 60 Co irradiation dose was 1.8 cGy, and anti-mouse CD122 monoclonal antibody was injected at 150 μg / mouse.

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Abstract

The invention discloses a method for constructing humanized Ph chromosome positive acute lymphocytic leukemia mouse model, and the method comprises the following steps: acquisition of leukemia cells, pretreatment of NOD/SCID mouse and implantation of leukemia cells, and implantation of leukemia cells. Specifically, the method comprises: 60Co irradiation of sub-lethal dose combined with a pretreatment with rat anti-mouse CD122 monoclonal antibody, and injection of Ph+ALL bone marrow mononuclear cells from knee joint marrow cavity of NOD/SCID mouse, thereby a humanized Ph+ALL mouse model is established with high molding efficiency and good repeatability; cell sorting is not needed, and the molding process is easy to be operated; the method provides an experiment platform for carrying out mankind Ph+ALL research on the level of living animals, and will promote deep research of domestic Ph+ALL.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for constructing a humanized Ph chromosome-positive acute lymphoblastic leukemia mouse model. The establishment of a disease model that simulates the characteristics of clinical Ph chromosome-positive acute lymphoblastic leukemia can provide a mouse in vivo research platform for the study of the pathogenesis, diagnosis and treatment methods, and new drug development of Ph chromosome-positive acute lymphoblastic leukemia. Background technique [0002] Ph chromosome, as the most common chromosomal abnormality in adult acute lymphoblastic leukemia (ALL), has become a sign of poor prognosis in ALL. In recent years, with the widespread application of tyrosine kinase inhibitors, Ph chromosome-positive acute lymphoblastic leukemia (English name Philadelphia chromosome-positive acute lymphoblastic leukemia, abbreviated as Ph + ALL) curative effect has been significantly improved, but...

Claims

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Application Information

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IPC IPC(8): C12N5/077A01K67/027
Inventor 黄晓军孔圆刘艳荣王亚哲常英军秦亚溱赖悦云江倩江浩
Owner PEOPLES HOSPITAL PEKING UNIV
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