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Method, transplant and culture medium for culturing chordoma organoid

A culture medium, chordoma technology, applied in the field of biomedicine, can solve the problems of long modeling time, insufficient patients, and low success rate of modeling chordoma xenograft animal models

Active Publication Date: 2020-12-08
北京科途医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Innovative treatments for rare cancers often have insufficient patients to establish clinically beneficial trials
[0003] Chordoma is a low-grade malignant tumor with aggressive growth, and the tumor cells have the characteristics of slow and invasive growth, which leads to low success rate and long modeling time of chordoma xenograft animal model

Method used

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  • Method, transplant and culture medium for culturing chordoma organoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] This example is used to illustrate the preparation of chordoma organoids.

[0056] (1) Preservation and transportation of chordoma tissue samples

[0057] Surgical tissues from the sacrum of patients with clinically confirmed chordoma were obtained as chordoma tissue samples, then preserved in 0.9% sterile saline, placed on ice, and transported to the operating room within 48 hours.

[0058] (2) Acquisition of chordoma tissue cells

[0059] Place the chordoma tissue sample obtained above in a sterile petri dish, rinse twice with sterile PBS, and add an enzymatic hydrolysis solution after washing, wherein the enzymatic hydrolysis solution contains 10% FBS, 1.5 mg / mL I Type collagenase, 100U / ml of penicillin, 100mg / ml of streptomycin and the rest of DMEM medium, 0.1mL of the above enzymatic hydrolysis solution was added for every 1mg of chordoma tissue sample. Chordoma tissue samples were cut into 1mm 3 The left and right small pieces were then enzymatically hydrolyzed...

Embodiment 2

[0067] This example is used to illustrate the preparation of the graft of the present disclosure.

[0068] Continue culturing the chordoma organoids cultured in Example 1 until the diameter of a single chordoma organoid is not less than 1.0 mm, and then place the organoids in each culture well together with a surrounding coating layer with a thickness of not less than 1.0 mm Take it out to obtain the graft A of this embodiment.

Embodiment 3

[0070] The method of Example 1 was used to culture chordoma organoids, except that the concentration of epirubicin in the first medium was 25 mg / L, and the concentration of epirubicin in the second medium was 10 mg / L.

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Abstract

The disclosure relates to a method for culturing a chordoma organoid. The method comprises the steps: mixing chordoma tissue cells with a first culture medium and matrix glue, so as to obtain a firstpremix, wherein the first culture medium contains epirubicin, and the epirubicin has a concentration of 15mg / L to 25mg / L by taking the first culture medium as a reference; culturing the first premix for 3 to 7 days, so as to obtain a first culture; acquiring a liquid part from the first culture, and separating cell precipitate from the liquid part, wherein the cell precipitate contains chordoma cells; mixing the cell precipitate with a second culture medium and matrix glue, so as to obtain a second premix, wherein the second culture medium contains the epirubicin, and the epirubicin has a concentration of 5mg / L to 10mg / L by taking the second culture medium as a reference; and subjecting the second premix to culture amplification, thereby obtaining the chordoma organoid.

Description

technical field [0001] The present disclosure relates to the technical field of biomedicine, in particular to a method for culturing chordoma organoids, a transplant for constructing a chordoma xenograft model and its application, and a method for producing a chordoma xenograft mouse model, and a medium for chordoma organoid culture. Background technique [0002] Chordoma is a rare congenital primary bone tumor with an incidence of less than 1 / 1 million. Chordoma is extremely insensitive to traditional radiotherapy and chemotherapy, and it is difficult to completely remove it by surgery. The recurrence rate is 50% for skull base chordomas and 30-40% for spinal chordomas. At present, the treatment of chordoma has become a major challenging clinical problem in the field of spinal tumor surgery and neurosurgery. Innovative treatments for rare cancers often have insufficient patients to establish clinically beneficial trials. [0003] Chordoma is a low-grade malignant tumor ...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/077A01K67/027
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331C12N5/0654C12N5/0693C12N2501/01C12N2501/11C12N2501/33C12N2501/39C12N2501/999C12N2533/90
Inventor 孙志坚李程肖金平康平
Owner 北京科途医学科技有限公司
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