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Separate culture method of primary tumor cells

A primary tumor cell separation and culture technology, applied in cell dissociation methods, tumor/cancer cells, culture process, etc., can solve the problem of low culture success rate and clinical consistency rate, inability to fully disperse tumor cells, primary tumor cell Long cultivation period and other problems, to achieve the effect of easy to adhere to the wall and improve the survival rate

Inactive Publication Date: 2019-03-01
钱成穗
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] It has always been limited by the technology of in vitro culture of primary tumor cells. When dispersing tumor cells, the tumor cells in the selected sample tissue cannot be fully dispersed; moreover, the culture period of the cultured primary tumor cells is long, The culture success rate and clinical consistency rate are relatively low

Method used

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  • Separate culture method of primary tumor cells
  • Separate culture method of primary tumor cells
  • Separate culture method of primary tumor cells

Examples

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Embodiment 1

[0042] After obtaining informed consent from the patient, tumor samples and normal samples from a patient undergoing lung cancer surgery were obtained from a cancer hospital in Beijing. The patient was male, aged 85, and clinically diagnosed as stage IV lung cancer.

[0043] S1. Place the fresh lung cancer tumor tissue in a petri dish, and add an appropriate amount of PBS to it, use ophthalmic forceps and ophthalmic scissors to remove blood (blood clot), fat, necrotic tissue and connective tissue on the tissue, and retain the area rich in lung cancer tumor cells , and washed 3 times with PBS containing antibiotics;

[0044] S2. Put the lung cancer tissue into a new petri dish and add a small amount of DMEM medium to it, use ophthalmic scissors to cut the tissue into pieces, and transfer it to a 15ml centrifuge tube, wash it with PBS for 3 times, and the tissue pieces will be removed automatically. After sinking, remove PBS;

[0045] S3. Transfer the tissue pieces into a cultu...

Embodiment 2

[0051] After obtaining informed consent from the patient, tumor samples and normal samples from a patient undergoing breast cancer surgery were obtained from a cancer hospital in Beijing. The patient was female, aged 52, and clinically diagnosed as Her2-positive breast cancer stage III.

[0052] S1. Place fresh breast cancer tumor tissue in a petri dish, and add an appropriate amount of PBS to it, use ophthalmic forceps and ophthalmic scissors to remove blood (blood clot), fat, necrotic tissue and connective tissue on the tissue, and retain abundant breast cancer tumor cells area, and washed 3 times with PBS containing antibiotics;

[0053] S2. Put the breast cancer tissue into a new petri dish and add a small amount of DMEM medium to it, use ophthalmic scissors to cut the tissue into pieces, and transfer it to a 15ml centrifuge tube, wash it with PBS 3 times, and the tissue pieces After automatic sinking, remove PBS;

[0054] S3. Transfer the tissue pieces into a culture bot...

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Abstract

The invention relates to the technical field of cell separation, in particular to a separate culture method of primary tumor cells. A composite enzyme solution is adopted for decomposing cancer tissue, wherein collagenase, trypsin and hyaluronidase in the composite enzyme solution mutually cooperate, looseness of bridging structures among the cells can be accelerated, the cancer tissue is converted to be flocculent from being blocky, thus, the cells in cell aggregate are evenly dispersed in cell suspension, and the numbers of cell alliances and single cells suspended in cell sap are increased;after inoculated culture is completed, the cells easily adhere to walls to grow, and therefore the survival rate of the cells is increased; serum extracted from the body of a patient suffering from tumors is added into a culture medium, a microenvironment most suitable for the growth of the cancer cells is provided for the cancer cells, the culture period is shortened, the culture success rate is80% or above, and the clinic consistency rate is 95% or above.

Description

technical field [0001] The invention relates to the technical field of cell separation, in particular to a primary tumor cell separation and culture method. Background technique [0002] Tumor cells are tumors in essence. Cancer is a genetic disease, but not hereditary. It refers to the cells under the action of tumorigenic factors, the gene has changed, and the normal regulation of its growth is lost, resulting in abnormal proliferation. Can be divided into two categories of benign and malignant tumors. The former grows slowly, has a clear boundary with surrounding tissues, does not metastasize, and is not harmful to human health. [0003] The process of tumorigenesis includes initiating mutation, latency, carcinogenesis, and evolution. The initial mutation refers to the gene mutation of the cells under the action of carcinogens, but if there is no appropriate environment after the mutation, it will not develop into a tumor. This stage is called the incubation period; ...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/02C12N2509/00
Inventor 钱成穗
Owner 钱成穗
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