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Transplant for constructing cholangiocarcinoma xenograft model and its preparation method and application

A technology for xenotransplantation and cholangiocarcinoma, which is applied in the field of biomedicine, can solve the problems of long time required, limited amount of tissue, low success rate of cholangiocarcinoma xenotransplantation model, etc., and achieves the effect of short time required and high success rate

Active Publication Date: 2022-04-19
科途(固安)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] However, when using the primary or metastatic tumor tissue blocks of patients with cholangiocarcinoma as transplants, the amount of tissue obtained is limited, the success rate of establishing cholangiocarcinoma xenograft models is low, and it takes a long time

Method used

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  • Transplant for constructing cholangiocarcinoma xenograft model and its preparation method and application

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preparation example Construction

[0038]According to the present disclosure, the enzymatically hydrolyzed cholangiocarcinoma tissue used in the present disclosure can be obtained from cholangiocarcinoma tissue through enzymatic hydrolysis. Exemplarily, the preparation method of the enzymatically hydrolyzed cholangiocarcinoma tissue may include, for example: (1) mixing fragments of cholangiocarcinoma tissue with an enzymolysis solution and performing enzymolysis to obtain an enzymolysis material, wherein the cholangiocarcinoma Tissues include primary cholangiocarcinoma tissue and / or metastatic cholangiocarcinoma tissue; the enzymolysis solution contains RPMI 1640 medium, 2-5% fetal bovine serum albumin, and 1-3 mg / mL collagen hydrolytic enzyme; Relative to 1 part by weight of the cholangiocarcinoma tissue fragments, the amount of the enzymatic hydrolysis solution is 5 to 10 parts by volume; (2) centrifuging the enzymatic hydrolysis material, collecting the precipitate, and obtaining the cholangiocarcinoma after ...

Embodiment 1

[0050] This example is used to illustrate the preparation method of the graft of the present disclosure.

[0051] (1) Preservation of cholangiocarcinoma tissue

[0052] The fresh surgical tissue of the primary tumor of cholangiocarcinoma was preserved in the above-mentioned cholangiocarcinoma culture medium, and transported to the operating room within 48 hours of cold chain at 4 degrees.

[0053] (2) Preparation of cholangiocarcinoma tissue after enzymatic hydrolysis

[0054] The culture medium was removed, and the cholangiocarcinoma tissue was placed in a 6 cm Petri dish. Configure RPMI 1640 medium containing 2% FBS and 2 mg / mL collagen hydrolytic enzyme as the enzymolysis solution. After the enzymolysis solution is filtered and sterilized and warmed at 37°C, it is added to the culture dish with cholangiocarcinoma tissue, of which, 1g For cholangiocarcinoma tissue, add 8 mL of enzymatic hydrolysis solution. Use sterilized scissors to cut the cholangiocarcinoma tissue into...

Embodiment 2

[0059] The graft B of the present disclosure was prepared by the method of Example 1, except that in this example, the tissue block of the primary cholangiocarcinoma tumor in Example 1 was replaced with the tissue block of the metastatic cholangiocarcinoma from the same patient.

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Abstract

The present disclosure relates to a graft for constructing a cholangiocarcinoma xenograft model and its preparation method and application. The preparation method comprises: a. mixing Matrigel and cholangiocarcinoma medium to obtain a cholangiocarcinoma medium-Matrigel mixture; b. . Mixing the enzymatically hydrolyzed cholangiocarcinoma tissue with the cholangiocarcinoma medium-matrigel mixture, culturing and expanding, to obtain a transplant containing cholangiocarcinoma organoids. In the above method, the enzymatically hydrolyzed cholangiocarcinoma tissue is used to culture the transplanted body containing the cholangiocarcinoma organoid, and the success rate of constructing the cholangiocarcinoma xenograft model using the transplanted body is relatively high and the required time is relatively short.

Description

technical field [0001] The present disclosure relates to the technical field of biomedicine, in particular to a graft for constructing a cholangiocarcinoma xenograft model, a preparation method and application thereof. Background technique [0002] Cholangiocarcinoma refers to malignant tumors originating from the extrahepatic bile duct, including the bile duct from the hilar region to the lower end of the common bile duct. Its etiology may be related to diseases such as bile duct stones and primary sclerosing cholangitis. Surgical treatment, radiotherapy, chemotherapy and other methods can be used clinically, but the effect is not obvious and the prognosis is poor. In order to improve the therapeutic effect of cholangiocarcinoma, in-depth research on the pathogenic mechanism and drug resistance of cholangiocarcinoma is needed, and the animal xenograft model is an important tool for research. [0003] In the related art, the primary or metastatic tumor tissue block of a pa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/02C12N5/09
CPCA01K67/02C12N5/0693C12N2500/38C12N2533/90
Inventor 孙志坚康平李程肖金平
Owner 科途(固安)生物科技有限公司
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