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Culture medium, culture method and detection method of thyroid cancer organoid

A technology of thyroid cancer and culture medium, applied in the field of culture medium of thyroid cancer organoids, can solve the problem that the thyroid cancer organoid model culture steps, the optimal culture medium formulation and identification method are not specified, and it is difficult to obtain a thyroid cancer organoid model And other issues

Pending Publication Date: 2021-05-25
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is only one study on thyroid cancer organoids reported at home and abroad (Sondorp et al., Cancers (Basel), 2020, 12(11): 3212), but it is difficult to obtain long-term stable cultured thyroid cancer organoids in vitro model, and the culture steps, optimal medium formula and identification methods of the thyroid cancer organoid model were not specifically clarified

Method used

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  • Culture medium, culture method and detection method of thyroid cancer organoid
  • Culture medium, culture method and detection method of thyroid cancer organoid
  • Culture medium, culture method and detection method of thyroid cancer organoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1: culture medium

[0074] A culture medium for thyroid cancer organoids, the medium is composed of Advanced DMEM / F12 medium and the following components: B-27, N-acetylcysteine, nicotinamide, R-spondin 1 recombinant protein, noggin , epidermal growth factor, SB202190, A83-01, fibroblast growth factor-7, fibroblast growth factor-10, thyrotropin and Y-27632; in the medium, the total volume of the medium Calculated, the volume percentage of the B-27 is 1%; the concentration of the N-acetylcysteine ​​is 1.25mM; the concentration of the nicotinamide is 10mM; the concentration of the R-spondin 1 recombinant protein is 500ng / mL; the concentration of the noggin is 100ng / mL; the concentration of the epidermal growth factor is 50ng / mL; the concentration of the SB202190 is 10μM; the concentration of the A83-01 is 500nM; the fibroblast The concentration of growth factor-7 is 5 ng / mL; the concentration of the fibroblast growth factor-10 is 5 ng / mL; the concentration of th...

Embodiment 2

[0075] Example 2: culture medium

[0076] A culture medium for thyroid cancer organoids, the medium is composed of Advanced DMEM / F12 medium and the following components: B-27, N-acetylcysteine, nicotinamide, R-spondin 1 recombinant protein, noggin , epidermal growth factor, SB202190 and thyrotropin; in the medium, based on the total volume of the medium, the volume percentage of the B-27 is 1%; the N-acetylcysteine The concentration is 1.25mM; the concentration of the nicotinamide is 10mM; the concentration of the R-spondin 1 recombinant protein is 500ng / mL; the concentration of the noggin is 100ng / mL; the concentration of the epidermal growth factor is 50ng / mL; the concentration of the SB202190 is 10 μM; the concentration of the thyroid-stimulating hormone is 10 μg / mL; the balance is Advanced DMEM / F12 medium.

Embodiment 3

[0077] Example 3: Culture of Thyroid Cancer Organoids

[0078] Operation: Take the culture medium described in Example 1 or Example 2, respectively, and take thyroid cancer tissues from 8 different patients obtained clinically, and culture thyroid cancer organoids according to the following steps:

[0079] (1) Take the thyroid cancer tissue, wash it, and cut it into pieces of about 1mm 3 Digested with type II collagenase at 37°C for 1 hour;

[0080] (2) Add Hanks solution to stop the digestion; centrifuge, discard the supernatant to obtain the first precipitate, resuspend the first precipitate with Hanks solution, filter with a 70 μm cell strainer, count the cells, and discard the supernatant Supernatant liquid, get the second precipitate, mix with the second precipitate with DMEM / F12 medium, add Matrigel and resuspend, the volume ratio of described Matrigel and DMEM / F12 medium is 3:1, inoculate, After incubation for 2 minutes, invert for another 8 minutes to solidify the Ma...

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Abstract

The invention provides a culture medium, a culture method and a detection method of a thyroid cancer organoid, the culture medium comprises a basic culture medium, B-27, N-acetylcysteine, nicotinamide, R-spondin 1 recombinant protein and other components, the culture medium has high thyroid cancer organoid culture success rate, high passage frequency, and good passage stability after repeated cryopreservation and resuscitation; the thyroid cancer organoid obtained by adopting the culture medium and the culture method is high in reducibility; and the detection method is comprehensive and accurate.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a culture medium, a culture method and a detection method of thyroid cancer organoids. Background technique [0002] Thyroid cancer is a common endocrine malignancy. my country is a high-incidence area, and its incidence is increasing year by year. According to tumor origin and differentiation, thyroid cancer can be divided into: papillary thyroid cancer (PTC), follicular thyroid cancer (FTC), medullary thyroid cancer (Medullary thyroid cancer, MTC) and Anaplastic thyroid cancer (ATC). Among them, PTC accounts for about 85%-90% of all thyroid cancers. Although most differentiated thyroid cancers can be remitted after surgery, postoperative radioactive iodine-131, and thyroid hormone suppression therapy, some patients are refractory to radioiodine in their natural history or after treatment. The survival rate of patients with radioactive iodine refractory, metastatic or adv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/6858G01N1/30
CPCC12N5/0693C12Q1/6858G01N1/30C12N2509/00C12N2509/10C12N2501/50C12N2500/32C12N2500/38C12N2501/415C12N2501/998C12N2501/11C12N2501/405C12N2501/375C12N2501/117C12N2501/119C12N2501/15C12N2513/00C12Q2535/122
Inventor 黄卫人陈栋
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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