Method for constructing ovarian cancer transplantation tumor model based on organoid method and application of method

A technology of ovarian cancer and organoids, which is applied in the field of medicine, can solve the problems of drug sensitivity, difficult to raise, and long tumor formation cycle of patients who urgently need drug treatment in clinical practice

Inactive Publication Date: 2019-03-01
上海美峰生物技术有限公司
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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, compound ovarian cancer cells derived from patients to cultivate cell-3D material complexes, and then directly inoculate the cell-3D material complexes on Normal immunized mice were subcutaneously constructed in vivo patient-derived ovarian cancer xenograft tumor models, thereby solving the existing human ovarian cancer xenograft tumor models using nude mice, SCID mice and other immunodeficient mice that are not easy to raise, expensive, and have a low tumor formation rate In particular, it solves the problem that the immunodeficient mouse human ovarian cancer xenograft model cannot reflect the important role played by the body's immune system in the occurrence and development of tumors, and the immunodeficient mouse human xenograft tumor model has a tumor formation cycle Long (usually 3-5 months), it is difficult to solve the drug sensitivity needs of patients who urgently need drug treatment in clinical practice

Method used

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  • Method for constructing ovarian cancer transplantation tumor model based on organoid method and application of method
  • Method for constructing ovarian cancer transplantation tumor model based on organoid method and application of method
  • Method for constructing ovarian cancer transplantation tumor model based on organoid method and application of method

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specific Embodiment approach 1

[0020] Specific embodiment one: The organoid-based method of this embodiment constructs an ovarian cancer xenograft tumor model, which is carried out according to the following steps:

[0021] 1. Obtain human primary ovarian cancer tumor cells: take ovarian cancer cells and ovarian cancer tissue mesenchymal cells, and use DMEM containing 10% fetal bovine serum, 1% penicillin by mass percentage and 1% chain by mass percentage Mix the above cells in the complete culture solution of mycin and count them with trypan blue; if the number of live cells counted by trypan blue is >95%, adjust the cell concentration to 2×10 7 / mL, inoculated into the culture plate or culture bottle and placed in 37℃, 5% CO 2 Incubator, get human primary ovarian cancer cell suspension;

[0022] 2. After soaking the microcarrier in 75% alcohol by volume for 24 hours, wash it 4 times with 1×PBS buffer solution, place it in DMEM medium and incubate it for 24~72 hours; The cell suspension was mixed with th...

specific Embodiment approach 2

[0024] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the microcarrier is composed of a multi-layer hole-like strip-like cross-linked and curled organic composite polymer that can be positively charged, containing "labyrinth" "like irregular structure. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0025] Embodiment 3: This embodiment differs from Embodiment 1 in that: the volume ratio of the human primary ovarian cancer cell suspension obtained in Step 1 to the incubated microcarrier is 3-4:1. Others are the same as in the first embodiment.

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Abstract

The invention discloses a method for constructing an ovarian cancer transplantation tumor model based on an organoid method, and belongs to the field of medicines. According to the invention, an "organoid" culture technology is adopted, and a micro-carrier is used as a support material for culturing a patient-derived ovarian cancer cell-3D material complex; then the cell-3D material complex is directly inoculated under the skin of a normal immune mouse, and a patient-derived ovarian cancer transplantation tumor model is constructed, so that a problem that immune-deficiency mice such as severecombined immunodeficiency (SCID) mice, nude mice and the like are adopted in existing human ovarian cancer transplantation tumor models, the price is high, breeding is difficult, the tumor forming rate is low and the like can be solved, especially a problem that the existing immune-deficiency mouse human ovarian cancer transplantation tumor model cannot reflect impotent effects of an organism immune system in occurrence and development processes of tumors is solved, and a problem that an immune-deficiency mouse human transplantation tumor model is long in tumor forming period and cannot meet drug allergy requirements of patients urgently needing clinic drug therapy is solved. The method is applied to the field of mouse model construction.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to a medical tumor animal model-a patient-derived normal immune mouse ovarian 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. Applications. Background technique [0002] Tumor is a frequently-occurring and common disease that seriously threatens human health, and its pathogenesis has not been fully elucidated. Ovarian cancer is one of the most common malignant tumors. According to the statistics of the American Cancer Society in 2018, the death rate of ovarian cancer still ranks first among female reproductive system tumors. Therefore, it is of great significance to further explore the mechanism of occurrence and development of ovarian tumors and more effective treatment methods through experimental animal models. [0003] New research results, organoids were rated ...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12N5/09C12Q1/02
CPCA01K67/0271A01K2227/105A01K2267/0331C12N5/0693C12N2531/00G01N33/5011G01N33/5044
Inventor 洪丰彭存旭毕研贞潘红娜黄蓉王全全张凯谌通张凤梅上舒振峰
Owner 上海美峰生物技术有限公司
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