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High-throughput screening method for anti-breast cancer drugs

A screening method and anti-breast cancer technology, applied in the biological field, can solve problems such as complex detection, and achieve the effects of improving growth rate, less side effects, and improving survival rate

Active Publication Date: 2018-12-14
爱克精医(北京)生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method involves the gene level, and the detection is more complicated

Method used

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  • High-throughput screening method for anti-breast cancer drugs
  • High-throughput screening method for anti-breast cancer drugs
  • High-throughput screening method for anti-breast cancer drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 A high-throughput screening method for anti-breast cancer drugs

[0058] Specifically include the following steps:

[0059] (1) Obtain breast cancer cells: Remove the adipose tissue of the obtained breast cancer cells under aseptic conditions, cut into 1mm size, and then digest with 4mL digestive enzyme for 2 hours, each ml of digestive enzyme contains 225 units of collagenase / transparent Protonidase and 13 units of dispase, and then stop the digestion with DMEM cell culture medium containing 10% calf serum that is 4 times the volume of the breast cancer tumor tissue, collect the digestive juice, collect the precipitate after centrifugation, and obtain dispersed breast cancer cells;

[0060] (2) Expansion culture of breast cancer cells: The resuspended pellet of breast cancer cells obtained in step (1) was subjected to fidelity expansion culture in the laboratory, placed in a culture bottle and placed in 37°C CO 2 Cultured in an incubator, cultured for 3 week...

Embodiment 2

[0069] Example 2 A high-throughput screening method for anti-breast cancer drugs

[0070] The specific operation steps are:

[0071] (1) Obtain breast cancer cells: remove the adipose tissue of the obtained breast cancer cells under aseptic conditions, cut into 2mm in size, and then digest with 5mL of digestive enzymes for 3 hours, each ml of digestive enzymes contains 225 units of collagenase / transparent Protonidase and 13 units of dispase, and then stop the digestion with DMEM cell culture medium containing 10% calf serum that is 4 times the volume of the breast cancer tumor tissue, collect the digestive juice, collect the precipitate after centrifugation, and obtain dispersed breast cancer cells;

[0072] (2) Expansion culture of breast cancer cells: The resuspended pellet of breast cancer cells obtained in step (1) was subjected to fidelity expansion culture in the laboratory, placed in a culture bottle and placed in 37°C CO 2 Cultured in an incubator, cultured for 2 week...

Embodiment 3

[0081] Embodiment 3 A kind of high-throughput screening method of anti-breast cancer drug

[0082] The specific operation steps are:

[0083] (1) Obtain breast cancer cells: remove the adipose tissue of the obtained breast cancer cells under aseptic conditions, cut into 2mm in size, and then digest with 5mL of digestive enzymes for 2.5 hours, each ml of digestive enzymes contains 225 units of collagenase / transparent Protonidase and 13 units of dispase, and then stop the digestion with DMEM cell culture medium containing 10% calf serum that is 4 times the volume of the breast cancer tumor tissue, collect the digestive juice, collect the precipitate after centrifugation, and obtain dispersed breast cancer cells;

[0084] (2) Cultivation of breast cancer cells: The resuspended pellet of breast cancer cells obtained in step (1) was cultured in the laboratory for fidelity amplification, placed in culture bottles and placed in 37°C CO 2 Cultured in an incubator, cultured for 2.5 we...

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Abstract

The invention provides a high-throughput screening method for anti-breast cancer drugs. The method is different from previous methods, and can provide accurate individualized treatment selection for cancer patients. The concentrations of amphotericin B and gentamicin in a tumor cell culture solution are controlled to improve the survival rate of tumor cells, effectively maintain the cell morphology and promote the absorption of nutrients by the tumor cells. The concentrations of insulin and a growth factor EGF in the tumor cell culture solution are controlled to improve the growth rate of thetumor cells and improve the cell activity, so a large number of highly active tumor cells can be obtained; and certain proportions of the insulin and the growth factor can be used to reduce the use amount of fetal calf serum, the fetal calf serum has a high price, so the reduction of the use amount facilitates resource saving; and the half-inhibitory concentration (IC50) error of the drug determined through drug screening using the cultured cells is small, so the detection frequency can be reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a high-throughput screening method for anti-breast cancer drugs. Background technique [0002] Breast cancer is the most common malignant tumor among Chinese women, with a high incidence rate, which seriously threatens women's health. Breast cancer is a systemic disease. Even in the early stage, a small amount of breast cancer cells are transferred to other organs of the body. According to statistics, the incidence rate accounts for 7-10% of various malignant tumors in the whole body. Refers to breast cancer patients with negative estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2), accounting for 15-20% of breast cancer, characterized by high recurrence , early metastasis and poor prognosis and other clinical characteristics, the current main treatment includes local treatment (surgery and radiotherapy) and systemic treatment (che...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12N5/09
CPCC12N5/0693C12N2500/12C12N2500/25C12N2501/11C12N2501/33C12N2501/39C12N2501/392C12N2501/727C12N2503/02G01N33/5011G01N2500/10
Inventor 马永贤
Owner 爱克精医(北京)生物医药科技有限公司
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