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Macaca mulatta hepatocellular carcinoma model, macaca mulatta hepatocellular carcinoma cell line and application of macaca mulatta hepatocellular carcinoma model and macaca mulatta hepatocellular carcinoma cell line

A technology of liver cancer cells and rhesus monkeys, applied to preparations for in vivo experiments, medical preparations containing active ingredients, organic active ingredients, etc.

Active Publication Date: 2016-02-24
WEST CHINA HOSPITAL SICHUAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no reproducible rhesus monkey liver cancer model and transplantable rhesus monkey hepatocellular carcinoma cell line in the world

Method used

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  • Macaca mulatta hepatocellular carcinoma model, macaca mulatta hepatocellular carcinoma cell line and application of macaca mulatta hepatocellular carcinoma model and macaca mulatta hepatocellular carcinoma cell line
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  • Macaca mulatta hepatocellular carcinoma model, macaca mulatta hepatocellular carcinoma cell line and application of macaca mulatta hepatocellular carcinoma model and macaca mulatta hepatocellular carcinoma cell line

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Experimental program
Comparison scheme
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Embodiment 1

[0037] The preparation of embodiment 1 animal model of the present invention

[0038] 1. Materials and methods:

[0039] 1.1 Experimental materials

[0040] Six healthy rhesus monkeys within one week of birth, male, weighing 0.5-0.8 kg, were purchased from Chengdu Greenhouse Biotechnology Co., Ltd.

[0041] 1.2 Main reagents

[0042] Diethylnitrosamine: purchased from Sigma Company.

[0043] Normal saline was purchased from Southwest Pharmaceutical Co., Ltd.).

[0044] 1.3 Preparation of the main solution

[0045] Preparation of diethylnitrosamine solution:

[0046] Take 25ml of DENA stock solution (Sigma), add 75ml of physiological saline for injection, and prepare 20% DENA aqueous solution, and store it in the dark for future use.

[0047] 1.4 Main Instruments

[0048] Disposable medical sterile syringes: purchased from Sichuan Shuanglu Medical Instrument Co., Ltd.

[0049] 1.5 Modeling method

[0050] Intraperitoneal injection of 20% DENA aqueous solution, 40 mg / kg...

Embodiment 2

[0076] Example 2 Establishment of rhesus monkey liver cancer cell line M12353

[0077] 1. Experimental method

[0078] 1. Rhesus macaque hepatocellular carcinoma tissue xenografting in nude mice

[0079] Take the liver cancer-bearing rhesus monkey M12353 that survived in Example 1, kill the liver cancer-bearing rhesus monkey M12353 under carbon dioxide inhalation anesthesia, and take liver tumor tissues for: (1) use 10% neutral formalin (2) Serum-free medium for xenotransplantation in nude mice; (3) Serum-free medium for cell culture.

[0080] 2. Establishment of cell lines

[0081] Take M12353 liver cancer tissue or transplanted tumor tissue from nude mice, remove hemorrhagic and necrotic tissue, trim it into 0.1-0.5mm tissue pieces, place the tissue pieces in a petri dish at a certain interval, and add HM culture medium (Hepatocyte medium) after standing for 30 minutes , Sciencell, 5201), according to the cell growth, change the medium every 1-2 days or use (Gibco, A111...

Embodiment 3

[0096] Example 3 Detection of tumorigenicity and growth characteristics of the rhesus liver cancer cell line M12353 of the present invention

[0097] 1. In vitro cultured rhesus monkey liver cancer cell line M12353 nude mice subcutaneously transplanted into tumors

[0098] Harvest the rhesus macaque liver cancer cell line M12353 cultured in vitro to the 50th generation with the preservation number of CCTCCNO: C2015142, resuspend the cell suspension with serum-free culture medium and Matrigel (1:1), and use the cell suspension to subcutaneously in nude mice Inoculation transplant, 1.5-5×10 6 Each inoculated animal, 2-4 points each time, repeated 5 times.

[0099] The results show that each point can form a tumor each time, and the tumor formation rate is 100%. The histopathological morphology of the tumor is as follows: Figure 12 As shown, the same pathological features as liver cancer tissue.

[0100] The experimental results show that the rhesus monkey liver cancer cell l...

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Abstract

The invention discloses a macaca mulatta hepatocellular carcinoma model preparation method. According to the method, diethylnitrosamine is applied to primates, and the application dosage of the diethylnitrosamine is 40mg / Kg / per time. The invention further provides a macaca mulatta hepatocellular carcinoma cell line and application thereof. A macaca mulatta hepatocellular carcinoma model can be used for evaluating hepatocellular carcinoma prevention factors, new anti-hepatocellular carcinoma medicine, hepatocellular carcinoma radiotherapy, hepatocellular carcinoma surgical treatment and novel technologies such as imaging diagnosis which cannot be completed by a rodent model.

Description

technical field [0001] The invention relates to a rhesus monkey liver cancer model, a rhesus monkey liver cancer cell line and application thereof. Background technique [0002] Liver cancer, Hepatocellular carcinoma, HCC, is a common malignant tumor of the liver, with 700,000 newly diagnosed cases every year. In 2012 alone, there were 782,000 new cases of liver cancer worldwide, and 746,000 deaths. Currently, because of the limited treatment options , Liver cancer is a serious threat to human health and life. At present, the incidence of liver cancer in mainland China has ranked first in the mortality rate of malignant tumors of the digestive tract, accounting for 55% of the global incidence and 45% of the global death toll. How to improve the diagnosis and treatment level of liver cancer and improve the quality of life of patients is the fundamental problem that needs to be solved urgently in the prevention and treatment of liver cancer. Liver cancer animal models, espec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K31/131
Inventor 王修杰钟治晖
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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