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Method for preparing ovarian cancer organoid

A technology for ovarian cancer cells and cancers, which is applied in the field of biomedicine and can solve the problem of difficult tissue blocks.

Active Publication Date: 2020-04-21
中关村科技租赁股份有限公司
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  • Abstract
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Problems solved by technology

[0003] In the prior art, large ovarian cancer tissue blocks obtained from ovarian cancer surgery are directly used for cultivation, and functional tests and drug screening tests for ovarian cancer are carried out. However, for patients with recurrence drug resistance or visceral metastasis, It is difficult to obtain large tissue blocks through surgery, so there is an urgent need to develop a method to grow ovarian cancer cell samples using a small amount of ovarian cancer tissue or cells

Method used

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  • Method for preparing ovarian cancer organoid

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Embodiment

[0040] 1. Preparation of feeder layer cells

[0041] Human endometrial stem cells (Beijing Crespo, CE20016) were plated at a density of about 5000 cells per well in a 96-well plate containing 50 μl of Cosmo thermosensitive hydrogel per well, and published on Stemedia TM WIT-I TM Incubate in the medium for 24 hours at 37°C. After the culture, the whole 96-well plate covered with human endometrial stem cells was placed in an X-ray irradiator for irradiation. The irradiation dose was 80KV, and the irradiation time was 30min. After the irradiation, the human uterus Intimal stem cells were used as feeder cells.

[0042] 2. Preservation of puncture tissue

[0043] Store fresh ovarian cancer puncture tissue samples in Stemedia containing penicillin 150U / ml and streptomycin 150ug / ml TM WIT-I TM culture medium, and transported to the laboratory at 4°C within 48 hours of cold chain.

[0044] 3. Ovarian cancer cell acquisition

[0045] Mince the ovarian cancer puncture tissue sa...

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Abstract

The invention relates to a method for preparing an ovarian cancer organoid, which is characterized by comprising the following steps of: a, mixing feeder layer cells, temperature-sensitive hydrogel, ovarian cancer cells and a first culture medium, and culturing for 7-9 days to obtain a first culture; wherein the feeder layer cells are human endometrial stem cells without a proliferation function;and b, changing the liquid of the first culture by using the first culture medium, and culturing until the ovarian cancer organoid is formed. According to the method disclosed by the invention, the ovarian cancer organoid matched with ovarian cancer tissue biological information can be cultured by utilizing a small amount of ovarian cancer tissues or cells, and the culture success rate is relatively high.

Description

technical field [0001] The present disclosure relates to the technical field of biomedicine, in particular to a method for preparing ovarian cancer organoids. Background technique [0002] Ovarian cancer is one of the most common cancers in female reproductive organs, and its incidence rate is second only to cervical cancer and uterine body cancer. Surgery combined with chemotherapy is the main treatment for ovarian cancer. Most patients are sensitive to chemotherapy after the first operation, but with the progress of chemotherapy, 50%-70% of patients gradually develop resistance to chemotherapy, resulting in ineffective treatment. In order to carry out functional tests and drug screening tests for ovarian cancer to further screen more drugs specifically targeting ovarian cancer cells, it is necessary to obtain ovarian cancer cell samples that match the biological information of ovarian cancer tissue. [0003] In the prior art, large ovarian cancer tissue blocks obtained fr...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/074
CPCC12N5/0682C12N5/0697C12N2501/727C12N2502/243C12N2502/30
Inventor 孙志坚康平李程黄思颖
Owner 中关村科技租赁股份有限公司
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