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Culture method of in-vitro substance used for scientific research experiments

A culture method and in vitro culture technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of high cost, low consistency, long culture time, etc., and achieve system stability, high success rate, Simple operation effect

Pending Publication Date: 2019-06-04
陕西茵莱生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] General organoid culture methods have low success rate, high cost, long culture time and low consistency for breast cancer organoid culture

Method used

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  • Culture method of in-vitro substance used for scientific research experiments
  • Culture method of in-vitro substance used for scientific research experiments
  • Culture method of in-vitro substance used for scientific research experiments

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] 1) Processing of patient-derived breast cancer tissue:

[0091] The sample tissue obtained from surgical resection of cancer tissue or biopsy from breast cancer patients was washed with PBS pre-cooled to 4°C. After the supernatant was clear, the sample was cut into 1 ~ 3mm 3 Add 5-15ml breast cancer organoid culture medium-1 containing 2mg / ml collagenase enzyme, incubate on a shaker at 37°C for 60-120 minutes, pipette up and down 30 times with a 10ml disposable plastic pipette Above, try to blow open the cell clumps. Filter with a 100-micron pore size filter to obtain primary breast cancer cells.

[0092] After centrifugation at 400rcf for 10 minutes, carefully remove the supernatant, add 10ml breast cancer organoid culture medium-1, pipette up and down 5-10 times, count the cells, and obtain breast cancer primary cells that can be used for organoid culture. Proceed to the next step of breast cancer organoid culture.

[0093] 2) In vitro culture of breast cancer pri...

Embodiment 2

[0099] 1) Processing of patient-derived breast cancer tissue:

[0100] The sample tissue obtained from surgical resection of cancer tissue or biopsy from breast cancer patients is washed with PBS pre-cooled to 4°C until the supernatant is clear, and the sample is cut into 3mm with tissue scissors in a 10cm petri dish 3 Add 20ml breast cancer organoid culture medium-1 containing 2mg / ml collagenase enzyme, incubate on a shaker at 37°C for 120 minutes, pipette up and down more than 30 times with a 10ml disposable plastic pipette, try to Cell clumps blow open. Filter with a 100-micron pore size filter to obtain primary breast cancer cells.

[0101] After centrifugation at 400rcf for 10 minutes, carefully remove the supernatant, add 10ml of breast cancer organoid culture medium-1, pipette up and down 10 times, count the cells, and obtain breast cancer primary cells that can be used for organoid culture to enter the next stage. One-step breast cancer organoid culture.

[0102] 2)...

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Abstract

The invention relates to the field of accurate medical treatment and cell biology, in particular to a culture method of an in-vitro substance used for scientific research experiments. The method comprises the following steps: 1, treatment of breast cancer tissues from patients: (1) treating fresh breast cancer sample tissues; (2) performing collagenase digestion after shearing; and (3) acquiring breast cancer primary cells; and 2, in-vitro culture of the breast cancer primary cells: (1) mixing the breast cancer primary cells with an organoid culture medium; (2) after solidification, adding thesolidified mixture into a breast cancer organoid culture solution for culture; and (3) acquiring a breast cancer organoid.

Description

technical field [0001] The invention relates to the fields of precision medicine and cell biology, in particular to a method for cultivating in vitro substances used in scientific research experiments. Background technique [0002] Breast cancer is the most common cancer among Chinese women. The incidence rate is 21.6 / 100,000 people. According to the data of "China Cancer Annual Report", there are 46,600 new cases of breast cancer every year. Although there are various treatment methods at present, including surgical treatment, targeted therapy and other treatment methods, which have achieved certain effects, there is still a certain blindness in the selection of treatment options and the prediction of clinical efficacy. [0003] The main reason is that breast cancer is a complex heterogeneous disease, which can be classified into different subtypes according to the expression of special genes, and there are significant differences in clinical manifestations, treatment prog...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
Inventor 扈晖郑维越
Owner 陕西茵莱生物科技有限公司
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