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Construction method and application of ovarian cancer organoid model

A construction method, ovarian cancer technology, applied in the direction of biochemical equipment and methods, instruments, scientific instruments, etc., to achieve the effect of avoiding the enrichment of tumor components and reducing the cost of culture

Pending Publication Date: 2022-06-10
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ovarian cancer organoid model has the disadvantages of high culture costs, and the immune components can only be retained for a short period of time, and tumor cells will be enriched during long-term culture.

Method used

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  • Construction method and application of ovarian cancer organoid model
  • Construction method and application of ovarian cancer organoid model
  • Construction method and application of ovarian cancer organoid model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] 1) Obtain ovarian cancer foci as ovarian cancer specimens, soak part of the specimens of cancer foci in sterile tubes filled with preservation solution, and place the sterile tubes on ice; the tissue preservation solution is formulated with high glucose The DMEM medium is based on adding Sijiqing, and the addition amount of the Sijiqing is 2-3% of the total volume of the high-sugar DMEM medium;

[0063] 2) In the aseptic operating table, wash the above-mentioned part of the cancer focus specimen in step 1) with tissue washing solution, cut off the necrotic tissue in the cancer focus, and select the cancer tissue with better activity and high degree of malignancy, and then The cancer foci were mechanically separated (shredded and scraped) with a sterile brush to obtain a homogenized suspension of tissue fragments; wherein, the formula of the tissue washing solution was based on high-sugar DMEM medium with the addition of Sijiqing and double antibodies, and Sijiqing The a...

Embodiment 2

[0077] 2) In the aseptic operating table, wash the above-mentioned part of the cancer focus specimen in step 1) with tissue washing solution, cut off the necrotic tissue in the cancer focus, and select the cancer tissue with better activity and high degree of malignancy, and then The cancer foci were mechanically separated (shredded and scraped) with a sterile brush to obtain a homogenized suspension of tissue fragments; wherein, the formula of the tissue washing solution was based on high-sugar DMEM medium with the addition of Sijiqing and double antibodies, and Sijiqing The amount of green added is 2-3% of the total volume of the high-sugar DMEM medium;

[0078] 3) After resuspending the suspension of tissue fragments with tissue washing solution, pass through a 70 μm filter, take cell spheroids smaller than 70 μm in size, and pass through a 40 μm filter upside down to obtain ovarian cancer organoid spheroids with a diameter of 40-70 μm;

[0079] 4) Add 5ml of split red solu...

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Abstract

The invention discloses a construction method and application of an ovarian cancer organ model, and the method comprises the following steps: firstly, obtaining an ovarian cancer specimen, soaking a part of the ovarian cancer specimen in a tissue preserving fluid, flushing the part of the ovarian cancer specimen with a tissue flushing fluid, and then cutting and scraping to obtain a homogenate tissue fragment suspension; re-suspending the filter screen, and cracking red to obtain a red-cracked ovarian cancer organ sphere; adding hydrogel into the ovarian cancer organoid spheres to obtain a mixed solution, and putting the culture plate inoculated with the cell spheres or the culture dish inoculated with the cell spheres into a cell incubator for culturing until the hydrogel is solidified; finally, the cell culture plate is taken out and placed in a sterile cell table, a corresponding organoid culture medium is added into the pore plate for culture, and the ovarian cancer organoid model is obtained.The hydrogel is used for replacing Matrigel, R-spondin, Noggin and L-wnt3A are all produced on cell supernatant, and the culture cost of existing ovarian cancer organoid is greatly reduced.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a method for constructing an ovarian cancer organoid model and its application. Background technique [0002] Ovarian cancer is currently the second leading cause of cancer-related death among gynecological malignancies, second only to cervical cancer. Since the early symptoms are not obvious and the onset is hidden, once diagnosed, most patients have entered the advanced stage. Moreover, there is no effective treatment for ovarian cancer, and the current 5-year survival rate is less than 50%. The standard treatment for ovarian cancer is debulking surgery plus chemoradiotherapy, and PARP inhibitors, as the first-line maintenance therapy for ovarian cancer, are approved for ovarian cancer patients with homologous recombination repair gene deletion or platinum-sensitive recurrence, but for PARP inhibition There is still a lack of effective treatment options for patients with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693G01N33/5011C12N2533/00
Inventor 孙朝阳陈刚刘陈黄雨涵陆富年秦天予张莉
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH