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Construction method and application of xenograft tumor models derived from different subtypes of Chinese breast cancer patients

A construction method and xenotransplantation technology, which are applied in the direction of medical raw materials derived from mammals, medical preparations containing active ingredients, and pharmaceutical formulas, can solve the problems of unrecorded success rate of primary PDX, irreversible changes in natural attributes, loss of Issues such as the characteristics of primary tumors, to achieve the effect of increasing the success rate of construction, stability and lasting stimulation

Active Publication Date: 2019-10-22
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the tumor model established by cell lines has many defects: because the continuously passaged tumor cell lines adapt to the environment of the culture dish and lack the tumor microenvironment such as non-tumor stromal cells, extracellular matrix, tumor microenvironmental factors, etc., eventually leading to tumor biological The biological behavior and natural properties of these cell lines are irreversibly changed, and the tumors formed after planting these cell lines into immunodeficient mice are homogeneous and lose the characteristics of the primary tumor, so they cannot objectively reflect the situation of the primary tumor
Chinese patent CN104546868A discloses a method for establishing a primary human breast cancer xenograft model based on the Chinese population, but the success rate of primary PDX constructed by this method has not been recorded in detail. After studying the success rate of primary model construction not satisfactory

Method used

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  • Construction method and application of xenograft tumor models derived from different subtypes of Chinese breast cancer patients
  • Construction method and application of xenograft tumor models derived from different subtypes of Chinese breast cancer patients
  • Construction method and application of xenograft tumor models derived from different subtypes of Chinese breast cancer patients

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1: Construction method described in the present invention (construct primary human breast cancer xenograft tumor model)

[0045] Take human breast cancer tissue specimens, put them into a sterile 4°C pre-cooled 15ml centrifuge tube filled with DMEM basal medium, keep them at 0-4°C, and process them within 1 hour.

[0046] In the ultra-clean workbench, put the above tumor tissue into a sterile petri dish, cut off the surrounding adipose tissue and necrotic tissue, and use a sterile scalpel to cut the tumor tissue P 0 Cut 3 After the small piece was removed, transfer it into a sterile 1.5ml EP tube, add 200ul each of calf serum and matrigel (BD company, product number 354234), and use a 1ml syringe to change 12 # Aspirate tumor tissue with a needle for use;

[0047] The 18-22g, 6-8 week old SPF grade NOD-SCID female mice were used for inoculation. The specific vaccination method: after the mice were anesthetized, the abdomen and the back of the neck of the mic...

Embodiment 2

[0048] Example 2: Primary transplanted tumor model breast cancer transplanted tumor passage

[0049] Waiting for Example 1 Primary tumor-bearing mouse tumor grows to 300-500mm 3 The tumor-bearing mice were selected to be sacrificed by carbon dioxide asphyxiation, and the body surface was sterilized. The breast cancer xenograft tumor tissue in the primary mouse model was dissected under a sterile environment, chopped and then digested with digestive juice (collagenase (Stemcell Co., Ltd.). , Cat. No. 07912) 1 part: M199 medium (Corning Company, Cat. No. R10-060-cv) 9 parts), digested in a water bath at 37°C for 1 hour, filtered to obtain a single-cell suspension, centrifuged at 350×g for 5 minutes to collect cells, and used After resuspending the cells with calf serum, count the number of living cells with trypan blue and adjust the cell concentration to 1×10 7 / ml, using a 1ml syringe to implant into the fourth pair of breast pads of SPF grade NOD-SCID female mice at the age ...

Embodiment 3

[0050] Example 3: Stable passage test of breast cancer tissues of different subtypes

[0051] The test was carried out with 12 cases of clinical human breast cancer tissues as specimens according to the construction method of Example 1 and the passaging method of Example 2 of the present invention, and the basic characteristics of breast cancer tissues between generations were counted, and the results are shown in Table 1.

[0052] Table 1 Stable passage test of different subtypes of breast cancer tissues

[0053]

[0054] Note: The number of passages listed in the table "Maintaining Stable Passage of Molecular Typing Markers" does not mean that it can only be passed to the listed number of passages, and it can be passed stably all the time. Passage data cannot be infinitely recorded in the table.

[0055] It can be seen from Table 1 that the successful primary xenograft tumor model constructed according to the construction method of the present invention can be stably pas...

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Abstract

The invention relates to the field of biomedical research and discloses a construction method and application of a model for xenograft tumors from different subtype Chinese patients with breast cancer. The construction method includes: breaking a human breast cancer tissue sample that is acquired, adding calf serum and matrix gelatin, and implanting to a fourth pair of mammary pads of the abdomen of a NOD-SCID female mouse; implanting an estrogen slow-release preparation subcutaneously in the back of the neck of the NOD-SCID female mouse; feeding the female mouse normally. The human breast cancer fresh tumor tissues are mated with the calf serum and matrix gelatin and are inoculated directly into the mammary pads of the NOD-SCID mouse, the primary human-derived breast cancer xenograft tumor model is established by subcutaneous implanting of the estrogen slow-release preparation, stable and lasting stimulation can be provided for the primary human-derived breast cancer xenograft tumors, and construction success rate for the primary model is improved at the premise of keeping basic characteristics of different generations of breast tumors consistent with those of clinical primary samples.

Description

technical field [0001] The invention relates to the field of biomedical basic research, in particular to a construction method and application of xenograft tumor models derived from different subtypes of Chinese breast cancer patients. Background technique [0002] Breast cancer is a common malignant cancer that seriously endangers women's physical and mental health, accounting for the second place among female cancers. Globally, more than 1.2 million women are estimated to be diagnosed with breast cancer each year. Although my country is a country with a relatively low incidence of breast cancer, due to the huge population base, the absolute number of breast cancer patients is large; what is more serious is that due to the three-fold increase in the incidence of female breast cancer in recent years, my country has become a breast cancer One of the fastest growing countries, breast cancer has become the number one killer of urban women. Therefore, it has become an urgent ta...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/027A61K35/13A61D7/00A61K31/565
Inventor 柳素玲何学艳杜锐凯
Owner UNIV OF SCI & TECH OF CHINA
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