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Method for establishing PDX (Patient Derived Xenograft) model of human blood tumor

A hematological tumor and a technology for establishing a method, which is applied in the field of establishing human hematological tumor PDX models, can solve the problems of insufficient time and quantity to obtain mice, unfavorable disease model establishment, and high experimental costs, etc., to achieve long life and improve The effect of high success rate and survival rate

Active Publication Date: 2019-03-19
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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  • Method for establishing PDX (Patient Derived Xenograft) model of human blood tumor
  • Method for establishing PDX (Patient Derived Xenograft) model of human blood tumor
  • Method for establishing PDX (Patient Derived Xenograft) model of human blood tumor

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Embodiment Construction

[0029] According to the following examples, the present invention can be better understood.

[0030] 1. Pretreatment of patient leukemia samples:

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

[0032] 2. ATG+complement incubation group: a part of the patient sample cells are added to ATG (200μg / head) and 1ml of the patient’s autologous serum (the serum contains complement, if the patient’s own complement is insufficient, a certain amount of rabbit complement can be supplemented), and the mixture is at 37 degrees Incubate for 1 hour. A part of the product is used for flow cytometry to identify the content of T lymphocytes. At the same time, the cells are resuspended and counted. 6 Each cell / 0.2ml mixture / mouse was inoculated into mice via tail vein i...

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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 constructing a blood tumor experimental animal model, in particular to a method for constructing a PDX model of human blood tumor. Background technique [0002] According to the latest statistics, the incidence of hematological tumors in my country is about 3-4 people per 100,000. Among the mortality rates of malignant tumors, it ranks 6th (male) and 8th (female), among children and adults under 35. China ranks first. The xenotransplantation (PDX) model of patient tumor tissue is a preclinical research and drug screening animal model for malignant tumors. This model directly transplants the patient's fresh tumor tissue into immunodeficient mice to establish a tumor tissue cell bank that can be passaged stably, providing important support for tumor-related basic and clinical research. But this model also has certain problems at this stage. On the one hand, the construction of PDX models requires the use of immunodefici...

Claims

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Application Information

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Patent Type & Authority Applications(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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