Ph+ acute B lymphocytic leukemia KQBL-84 cell line and construction method and application thereof

A technology of B lymphocytes and construction methods, applied in the fields of application, animal cells, tumor/cancer cells, etc., can solve the problems of high modeling cost, slow modeling speed, and low degree of malignancy, and achieve low modeling cost and high cost. The effect of fast modeling speed and high degree of malignancy

Inactive Publication Date: 2018-12-28
GUIZHOU EDUCATION UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, when using leukemia cell lines to construct animal models, there are often problems such as poor pertinence, low degree of malignancy, slow modeling speed and high modeling cost.

Method used

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  • Ph+ acute B lymphocytic leukemia KQBL-84 cell line and construction method and application thereof
  • Ph+ acute B lymphocytic leukemia KQBL-84 cell line and construction method and application thereof
  • Ph+ acute B lymphocytic leukemia KQBL-84 cell line and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (I) Collection of bone marrow cells from donor mice

[0038] 1) Select 6-week-old healthy C57bl / 6 mice with body weight between 24-26g as donor mice, and use CO 2 sacrifice the donor mouse;

[0039] 2) Disinfect the abdomen of the sacrificed donor mouse with 70% disinfectant alcohol in an ultra-clean bench;

[0040] 3) Take the femur, tibia, and hip bone on both sides of the donor mouse and put them in the pre-cooled 293T cell culture medium. The ratio of the 293T cell culture medium is DMEM+10%FBS+1% glutamine+1% double Anti-+1% non-essential amino acids;

[0041] 4) Use a 10ml syringe to absorb a sufficient amount of 293T medium, use a 27G1 / 2 needle to flush the bone marrow cells in the bone marrow cavity into a 50ml centrifuge tube, quickly and vigorously upside down to fully divide the bone marrow cells into single cells;

[0042] 5) Collect all the cells in a tube with a 10ml pipette, and filter with a 100μM sterile filter to remove the residue;

[0043] 6) Tak...

Embodiment 2

[0062] The KQBL-84 cell line obtained in Example 1 was used to rapidly construct a primary mouse tumor model.

[0063] Will 2×10 5 The KQBL-84 cells were injected into the recipient C57bl / 6 mice through the tail vein, and the recipient mice could show refractory / relapsed acute lymphoblastic leukemia phenotype within 8-12 days, and the recipient mice’s The detection results in peripheral blood (PB), spleen (spleen), and bone marrow (BM) were as follows: image 3 shown.

Embodiment 3

[0065] The target drug resistance test was performed on the recipient mice infected in Example 2.

[0066] Two comparison groups were set up, which were the targeted anti-tumor drug Dasatinib experimental group and the Vehicle blank reagent group for testing. Test results such as Figure 4 As shown, the results show that conventional tyrosine kinase inhibitor-targeted treatment of such mice is ineffective, as expected.

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Abstract

The invention discloses a Ph+ acute B lymphocytic leukemia KQBL-84 cell line and a construction method and an application thereof. The cell line is preserved in China Center for Type Culture Collection on July 25, 2018 with the preservation number of CCTCC NO: C2018164. The cell line is characterized in that an MSCV-P210 retrovirus carrying a human fusion gene BCR-ABL infects bone marrow cells ofa C57bl/6 donor mouse to obtain a primary human cell line stably expressing the BCR-ABL human protooncogene, and screening and culture are then performed to obtain a highly carcinogenic and stably inherited precursor B cell line. Cells of the line established in vitro are injected into a C57bl/6 homologous receptor mouse. The cell line can be used for efficiently constructing a primary B-ALL mousemodel. The model has higher malignancy, rapid disease modeling and low modeling cost, and is suitable for evaluation of treatment methods of refractory/relapsing B-ALL and pathogenesis study of patients with B-ALL in Philadelphia chromosome positive (Ph+).

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a disease model of Ph+ acute B lymphocytic leukemia and a construction method and application of the KQBL-84 cell line thereof. Background technique [0002] Acute B-cell lymphoid leukemia (B-ALL) is a common malignant hematological tumor originating from the abnormal proliferation of B lymphocytes in the bone marrow. The proportion is about 20%. A large number of abnormally proliferating naive B cells can inhibit normal hematopoietic function, and can also invade tissues outside the bone marrow, such as peripheral blood, spleen, lymph nodes, gonads, and liver. [0003] With the maturity of stem cell transplantation technology and the application of targeted drugs such as tyrosine kinase inhibitors (TKI), the treatment effect of acute lymphoblastic leukemia has changed greatly, and its remission rate and long-term survival rate have been significantly improved. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867
CPCC12N5/0694C12N15/86C07K14/47C12N2510/00C12N2740/10043
Inventor 潘聪陈艳胡以国李宁邱强
Owner GUIZHOU EDUCATION UNIV
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