A method for establishing human hematological tumor pdx model

A technology for establishing methods and models, applied in the fields of antibodies, cyclic peptide components, medical raw materials derived from mammals, etc., can solve the problems that the time and number of mice cannot be guaranteed, the establishment of unfavorable disease models, and the high experimental cost. Achieving the effect of long life, improved success rate and high survival rate

Active Publication Date: 2020-11-03
JIANGSU PROVINCIAL HOSPITAL OF TCM
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Problems solved by technology

However, many experimental institutions, including my country, do not have the independent intellectual property rights of these two types of mice, so they need to import them at high prices to conduct related experimental research. The cost of experiments is high, and the time and quantity of mice obtained cannot be guaranteed. Conducive to the establishment of large-scale disease models

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  • A method for establishing human hematological tumor pdx model
  • A method for establishing human hematological tumor pdx model
  • A method for establishing human hematological tumor pdx model

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Embodiment Construction

[0029] The present invention can be better understood from the following examples.

[0030] 1. Pretreatment of leukemia samples from patients:

[0031] 1. Collect 5-50ml of blood or 5-50ml of bone marrow from the patient, and use Ficoll separation medium for gradient centrifugation to separate the patient's mononuclear cells. After washing the mononuclear cells twice with normal saline, resuspend them with 0.5-2ml of normal saline, and the cells count.

[0032] 2. ATG+complement incubation group: Take a part of the patient sample cells and add ATG (200μg / body) and 1ml of the patient's own serum (serum contains complement, if the patient's own complement is insufficient, a certain amount of rabbit complement can be added), mix well and store at 37°C Incubate for 1 hour. A part of the product was retained for flow cytometric detection to identify the content of T lymphocytes, and the cells were resuspended and counted at the rate of 1×10 6 cells / 0.2ml mixture / mouse, and inocu...

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Abstract

The invention discloses a method for establishing a PDX (Patient Derived Xenograft) model of human blood tumor. The method comprises the steps of extracting a blood tumor cell of a patient, adding rabbit anti-human thymocyte immunoglobulin ATG (Anti-Thymocyte Globulin) and patient autologous serum, mixing, incubating and after completing the incubation, re-suspending the obtained cell and inoculating in a mouse; and feeding the inoculated mouse with CsA (Cyclosporine A) and lasting for 5-10 days starting from 2 days before the inoculation of the blood tumor cell. The method disclosed by the invention has the beneficial effects that a novel highly immunodeficient NCG mouse independently developed in China is firstly adopted to establish a human leukemia PDX model; by orally taking human thymocyte immunoglobulin pretreatment sample combined with the cyclosporine for a long term, donor-derived T cells are removed and inhibited; and by combining the toxic effect of the ATG on lymphocytes with the functional blocking effect of the CsA on T lymphocytes, the immune function of the T lymphocytes can be persistently inhibited and the success rate of PDX modeling of the blood tumor is significantly improved.

Description

technical field [0001] The invention relates to a method for establishing an experimental animal model of hematological tumors, in particular to a method for establishing a human hematological tumor PDX model. Background technique [0002] According to the latest statistics, the incidence rate of hematological tumors in my country is about 3-4 per 100,000, ranking 6th (male) and 8th (female) in the mortality rate of malignant tumors, and among children and adults under the age of 35. China ranks first. Patient tumor tissue xenograft (PDX) model is an animal model for preclinical research and drug screening created for malignancy. This model directly transplants fresh tumor tissue from patients into immune-deficient mice to establish a tumor tissue cell bank that can be stably passaged, providing important support for tumor-related basic and clinical research. However, there are still some problems with this model at this stage. On the one hand, the construction of PDX mode...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/13A61K39/395A61K35/13A61K35/16
CPCA61K35/13A61K35/16A61K38/13A61K39/395
Inventor 朱学军田芳姜鹏君吴坚孔祥图于慧于菊华徐祖琼代兴斌陈晓丽庞洁李峻张文曦倪海雯陈健一陈玉根
Owner JIANGSU PROVINCIAL HOSPITAL OF TCM
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