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Mouse model of acute lymphoblastic leukemia and modeling method

A technology of acute lymphoblastic leukemia and modeling method, applied in the field of acute lymphoblastic leukemia mouse model and modeling, can solve the problems of complex preparation process, and achieve the effects of improving uniformity, reducing accidental death, and stabilizing the model

Active Publication Date: 2017-03-22
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The first object of the present invention is to provide a modeling method of acute lymphoblastic leukemia mouse model to overcome the technical problems of complex preparation process in the prior art

Method used

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  • Mouse model of acute lymphoblastic leukemia and modeling method
  • Mouse model of acute lymphoblastic leukemia and modeling method
  • Mouse model of acute lymphoblastic leukemia and modeling method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Packaging of retroviruses with BCR-ABL P190:

[0067] Co-transfect 293FT cells with Mig190 and helper plasmids PGP and P-Eco using Lipofectamine 2000, collect the cell supernatant rich in Mig190 retroviral particles 72 hours after transfection, filter with a 0.45 μm filter and press Retro-concentin VirusPrecipitation Solution The instructions carry out the concentration of Mig190 virus, and the concentration factor is 100 times. The specific method of concentration of Mig190 virus is as follows:

[0068] Add the filtered supernatant to a 50mL centrifuge tube, 40mL / tube, then add Retro-concentin VirusPrecipitation Solution, 10mL / tube, mix well, let stand at 4°C for 24h, centrifuge at 1500g×2h×4°C, discard in a clean bench For the supernatant, add 500 μL / tube of 2% inactivated serum Opti-MEM medium and fully resuspend.

[0069] Aliquot 300 μL / tube of the concentrated Mig190 virus and store at -80°C.

[0070] Preparation of mouse hematopoietic cells:

[0071] BABL / c mic...

Embodiment 2

[0075] Packaging of retroviruses with BCR-ABL P190:

[0076] Co-transfect 293FT cells with Mig190 and helper plasmids PGP and P-Eco using Lipofectamine 2000, collect the cell supernatant rich in Mig190 retroviral particles 72 hours after transfection, filter with a 0.45 μm filter and press Retro-concentin VirusPrecipitation Solution The instructions carry out the concentration of Mig190 virus, and the concentration factor is 100 times. The specific method of concentration of Mig190 virus is as follows:

[0077] Add the filtered supernatant to a 50mL centrifuge tube, 40mL / tube, then add Retro-concentin VirusPrecipitation Solution, 10mL / tube, mix well, let stand at 4°C for 24h, centrifuge at 1500g×2h×4°C, discard in a clean bench For the supernatant, add 500 μL / tube of 2% inactivated serum Opti-MEM medium and fully resuspend.

[0078] Aliquot 300 μL / tube of the concentrated Mig190 virus and store at -80°C.

[0079] Preparation of mouse hematopoietic cells:

[0080] BABL / c mic...

Embodiment 3

[0084] Packaging of retroviruses with BCR-ABL P190:

[0085] Co-transfect 293FT cells with Mig190 and helper plasmids PGP and P-Eco using Lipofectamine 2000, collect the cell supernatant rich in Mig190 retroviral particles 72 hours after transfection, filter with a 0.45 μm filter and press Retro-concentin VirusPrecipitation Solution The instructions carry out the concentration of Mig190 virus, and the concentration factor is 100 times. The specific method of concentration of Mig190 virus is as follows:

[0086] Add the filtered supernatant to a 50mL centrifuge tube, 40mL / tube, then add Retro-concentin VirusPrecipitation Solution, 10mL / tube, mix well, let stand at 4°C for 24h, centrifuge at 1500g×2h×4°C, discard in a clean bench For the supernatant, add 500 μL / tube of 2% inactivated serum Opti-MEM medium and fully resuspend.

[0087] Aliquot 300 μL / tube of the concentrated Mig190 virus and store at -80°C.

[0088] Preparation of mouse hematopoietic cells:

[0089] BABL / c mic...

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Abstract

The invention provides a mouse model of acute lymphoblastic leukemia and a modeling method, and relates to the field of biotechnology. The preparation method comprises the following steps: infecting IL-7 stimulated hematopoietic cells with retroviruses with BCR-ABLP190; and transfusing the stimulated hematopoietic cells to syngenic mice. The method has the advantages of being simple and easy to implement in a modeling process and capable of establishing the ph+ALL mouse model in one step. According to the mouse model of the acute lymphoblastic leukemia provided by the invention, the sorted BCR-ABL+B cells are transfused to the syngenic mice for the second time without irradiation treatment, with performance consistent with that of mice of first episode; the model is good in stability; and in addition, the uniformity of recipient mice is enhanced and such risks as accidental death of the recipient mice due to irradiation are reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an acute lymphoblastic leukemia mouse model and a modeling method. Background technique [0002] Acute lymphoblastic leukemia (ALL) is a malignant tumor disease that originates from the abnormal proliferation of B or T lineage cells of lymphocytes in the bone marrow. Abnormally proliferating blast cells can gather in the bone marrow and inhibit normal hematopoietic function, and can also invade tissues outside the bone marrow, such as meninges, lymph nodes, gonads, and liver. my country has conducted a survey on the incidence of leukemia, and the incidence of ALL is about 0.67 / 100,000. The incidence rate in oil fields and polluted areas is significantly higher than the national incidence rate. ALL childhood (0 to 9 years old) is the peak incidence, which can account for more than 70% of childhood leukemia. ALL accounts for about 20 percent of adult leukemias in adults. [0003] A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/867C12N5/10A01K67/027
CPCA01K67/027A01K2227/105A01K2267/0331C12N5/0635C12N15/86C12N2740/10043
Inventor 陈翀徐开林祁娜王雪吴庆运曾令宇曹江闫志凌李振宇
Owner XUZHOU MEDICAL UNIV
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