Synovial sarcoma organoid culture method and medium, and transplant and use thereof

A technology for synovial sarcoma and culture medium, applied in the field of biomedicine, can solve the problems of limited diversity, small number of synovial sarcoma, and difficulty in constructing animal models of synovial sarcoma in large quantities, so as to promote in vitro growth and improve the success rate of culture. Effect

Active Publication Date: 2022-07-05
FUDAN UNIV SHANGHAI CANCER CENT +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the diversity of synovial sarcoma cell lines is limited, and the biological characteristics of synovial sarcoma tissue are quite different; the number of synovial sarcoma is small, and it is difficult to obtain, which makes it difficult to construct animal models of synovial sarcoma in large quantities, and Synovial sarcoma animal model has a long construction cycle, low tumor formation rate, and small tumor volume

Method used

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  • Synovial sarcoma organoid culture method and medium, and transplant and use thereof
  • Synovial sarcoma organoid culture method and medium, and transplant and use thereof
  • Synovial sarcoma organoid culture method and medium, and transplant and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] This example is used to illustrate the culture of synovial sarcoma organoids.

[0066] (1) Acquisition of synovial sarcoma tissue samples

[0067] The surgically excised or punctured tissues of patients with synovial sarcoma were used as synovial sarcoma tissue samples, which were stored in pre-cooled synovial sarcoma protective solution and transported to the operating room within 48 hours at 4°C.

[0068] (2) Acquisition of synovial sarcoma tissue cells

[0069] The synovial sarcoma tissue sample was placed in a sterile petri dish, washed twice with PBS at 4°C, and then an enzymatic hydrolysis solution was added, wherein 0.1 ml of the enzymatic hydrolysis solution was added to 1 mg of the synovial sarcoma tissue sample. Using sterile instruments, the synovial sarcoma tissue samples were cut into tissue blocks with a diameter of less than 1 mm, and then the culture dish was placed in an incubator at 37°C for enzymatic hydrolysis for 0.5 to 2 hours to obtain an enzymat...

Embodiment 2

[0079] This example is used to illustrate the application of synovial sarcoma organoids in the construction of PDX mouse model.

[0080] For the culture product obtained in Example 1, a synovial sarcoma organoid with a diameter of about 0.2 mm was taken out together with a matrigel layer within a thickness of 0.2 mm around it to obtain a transplant.

[0081] Mice (SPE grade female SCID mice, 4 weeks old, purchased from Beijing Weitong Lihua Laboratory Animal Co., Ltd.) were anesthetized with 0.1 ml / 20 g of sodium pentobarbital. After successful anesthesia, the mice were placed on a clean operating table, and the underarm skin of the mice was incised with sterile instruments, and a 3-5 mm deep incision was made on the muscle layer of the mice with ophthalmic forceps. Then gently pull up the muscle layer, and use ophthalmic forceps to pick up the treated graft and put it into the incision. The ophthalmic forceps were loosened, the muscle layer was smoothed, and the mouse skin w...

Embodiment 3

[0084] The synovial sarcoma organoids were cultured according to the method of Example 1, except that the concentration of valproic acid in the separation medium used in this example was 2.4 mmol / L, and the concentration of valproic acid in the expansion medium was 0.8 mmol / L.

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Abstract

The present disclosure relates to a method for culturing synovial sarcoma organoids, the method comprising: separating, amplifying and culturing synovial sarcoma tissue cells using a separation medium and an expansion medium in sequence to obtain an expanded culture product; The separation medium contains valproic acid, and based on the separation medium, the concentration of the valproic acid is 2 to 3 mmol / l, preferably 2.4 to 2.8 mmol / l; the expansion medium contains Valproic acid, based on the expansion medium, the concentration of the valproic acid is 0.5-1.5 mmol / l, preferably 0.8-1.2 mmol / l; the expansion culture product is separated and processed to obtain cell precipitation; sub-culturing the cell precipitation using the expansion medium to obtain a sub-culture product; obtaining the solid part of the sub-culture product to obtain a synovial sarcoma organoid.

Description

technical field [0001] The present disclosure relates to the technical field of biomedicine, in particular to a method for culturing synovial sarcoma organoids, a medium for culturing synovial sarcoma organoids, a graft for constructing a synovial sarcoma xenograft model and uses thereof , and a method for producing a synovial sarcoma xenograft mouse model. Background technique [0002] Synovial sarcoma is a malignant soft tissue tumor derived from synovial cells or mesenchymal cells differentiated into mesenchymal cells, accounting for about 7% to 10% of soft tissue tumors. According to reports, 90% of synovial sarcoma patients are younger than 50 years old, most are 15 to 35 years old, and most of them are male. At present, the main treatment for synovial sarcoma is surgery. However, about 50% of the patients have recurrence within 2 weeks after the operation, and about 40% of the patients have metastasis. Therefore, the surgical treatment of synovial sarcoma is not effec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A01K67/027
CPCC12N5/0693C12N5/0652A01K67/0271C12N2500/30C12N2501/11C12N2501/115A01K2207/12A01K2267/0331A01K2227/105
Inventor 黄稳定严望军李程肖金平程默蔡维泺孙志坚康平
Owner FUDAN UNIV SHANGHAI CANCER CENT
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