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Method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on organ-like method, and application thereof

A technology of immunizing mice and constructing methods, which is applied in the field of medicine, can solve problems such as drug sensitivity requirements of patients urgently in need of drug treatment, long tumor formation cycle, and difficulty in feeding

Inactive Publication Date: 2019-01-11
上海美峰生物技术有限公司
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  • Abstract
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Problems solved by technology

[0006] The purpose of the present invention is to use the "organoid" culture technology, using microcarriers (NMC2) as the scaffold material, and combining patient-derived liver cancer cells to culture the cell-3D material complex, and then directly inoculate the cell-3D material complex on normal Subcutaneously immunize mice to construct a patient-derived liver cancer xenograft tumor model in vivo, so as to solve the current human liver cancer xenograft tumor models that use nude mice, SCID mice and other immunodeficient mice, which are difficult to raise, expensive, and have a low tumor formation rate. ; especially to solve the problem that the immune-deficient mouse human liver cancer xenograft tumor model cannot reflect the important role played by the body's immune system in the occurrence and development of tumors, and the immunodeficiency mouse xenograft tumor model has a long tumor-forming cycle (generally It takes 3-5 months), it is difficult to solve the drug sensitivity needs of patients who urgently need drug treatment in clinical practice

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  • Method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on organ-like method, and application thereof
  • Method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on organ-like method, and application thereof
  • Method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on organ-like method, and application thereof

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specific Embodiment approach 1

[0021] Embodiment 1: The method for constructing the transplanted tumor model of liver cancer normal immune mice based on the organoid method of the present embodiment is carried out according to the following steps:

[0022] 1. Obtain human liver cancer tumor cells: take liver cancer cells and liver cancer tissue interstitial cells, and culture them completely in DMEM containing 10% fetal bovine serum, 1% penicillin and 1% streptomycin Mix the above cells evenly; if the number of viable cells counted by trypan blue is >95%, adjust the cell concentration to 2×10 7 / mL, then put into 37℃, 5% CO 2 incubator;

[0023] 2. Soak the microcarrier in 75% alcohol by volume for 24 hours, wash it 5 times with 1×PBS buffer solution, place it in DMEM medium and incubate it for 24-48 hours; the human primary liver cancer cells obtained in step 1 The suspension was mixed with the incubated microcarriers, and the temperature was 37°C, 5% CO 2 Cultivate in the incubator for 24-48h to obtain...

specific Embodiment approach 2

[0025] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the microcarrier is composed of a positively charged organic composite polymer, which is multi-layer hole-like cord-like, with enough cross-linked curls to form The "maze"-like irregular structure of space.

[0026] Others are the same as in the first embodiment.

[0027] The microcarrier pore size, surface positive charge density, and carrier particle size can be adjusted by chemical synthesis. It is a pure organic compound, not easy to pollute, does not contain impurities, and has the characteristics of low immunogenicity, biocompatibility, and metabolism. Provide a stable microenvironment for cell growth. Sufficient space inside the microcarrier solves the problem of uneven concentration of cell growth nutrients and metabolic waste. At the same time, because the microcarrier's "maze"-like irregular structure can act as a barrier (in a short period of time), it can block t...

specific Embodiment approach 3

[0029] Specific embodiment three: the difference between this embodiment and specific embodiment one is: the volume ratio of the human primary liver cancer cell suspension obtained in step one and the incubated microcarrier is 1:3~5, and the preferred value is 1: 4. Others are the same as in the first embodiment.

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Abstract

The invention belongs to the field of medicine, and relates to a method for constructing a patient-derived tumor xenograft model in mice immunized with hepatocellular carcinoma based on an organ-likemethod, and an application thereof. The aim of the invention is to use the organ-like culture technology, adopts microcarrier as scaffold material, and combines the liver cancer cells from patient toculture the cells. The invention also discloses an organ-like method for preparing the liver cancer cells-3D material complex, and then directly into that cell-3D material complex is inoculated subcutaneously into normal immunized mice to construct liver cancer transplanted tumor model from patients in vivo, which solved the problems of using nude mice, SCID mice and other immunodeficient mice, which are difficult to feed, expensive and lower tumor formation rate. Especially, it can solve the problem that human hepatocellular carcinoma transplanted tumor model in immunodeficiency mice can notreflect the important role of immune system in tumor occurrence and development, and human tumor transplanted tumor model in immunodeficiency mice has a long tumor formation cycle, which urgently needs the drug-sensitive demand of patients with clinical urgent need. The invention is applied to the field of mouse model construction.

Description

technical field [0001] The present invention belongs to the field of medicine, and in particular relates to a medical tumor animal model-a patient-derived normal immune mouse liver cancer xenograft model based on "organoids" and a method for establishing the model, as well as their application in the field of medicine and clinical practice. application. Background technique [0002] Tumor is a frequently-occurring and common disease that seriously threatens human health, and its pathogenesis has not been fully elucidated. Liver cancer is one of the most common malignant tumors, and its mortality rate ranks among the top three in the world. The 5-year survival rate after surgery is about 10%-20%. The treatment effect is unsatisfactory and the prognosis is poor. Therefore, establishing an animal model of liver cancer that can simulate clinical characteristics is of great significance for the research of liver cancer. [0003] New research results Organoids were rated as th...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/00A01K67/027
CPCA01K67/0271C12N5/0075C12N5/0693A01K2227/105A01K2267/0331
Inventor 洪丰毕研贞潘红娜王全全张凯谌通张凤梅
Owner 上海美峰生物技术有限公司
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