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Method for culturing lung tumor organoids

A technology for organoids and lung tumors, applied in the field of culturing lung tumor organoids, can solve the problems of difficult analysis and observation, delayed patient condition, and complicated operation, and achieve the effect of improving repeatability and stability, improving complicated operation, and simplifying the culture method.

Pending Publication Date: 2022-03-22
WEST CHINA HOSPITAL SICHUAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This culture method has disadvantages such as cumbersome operation, high cost, small scale, different organoid particle sizes, not in the same plane, difficult to analyze and observe, and cannot be mass-produced and used on a large scale.
At the same time, the existing methods for culturing tumor organoids have a long culture period, which may cause delays in the patient's condition

Method used

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  • Method for culturing lung tumor organoids
  • Method for culturing lung tumor organoids
  • Method for culturing lung tumor organoids

Examples

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preparation example Construction

[0042] (1) Preparation of lung decellularized ECM: The pig whole lung decellularized scaffold was prepared by Triton-SLES-Triton perfusion method. The obtained decellularized lung scaffold (pulmonary extracellular matrix) was further freeze-dried, crushed by a ball mill, and then enzymatically hydrolyzed with pepsin to obtain a lung decellularized extracellular matrix (ECM) gel.

[0043] (2) Using PDMS micropattern printing technology to prepare a patterned microarray master: using a laser to etch a silicon wafer with a specific pattern as a template, the etching accuracy is 1 μm, and the etching area is up to 12 inches, and polydimethylsiloxane is obtained by pouring the mold alkane (polydimethylsiloxane, PDMS) stamp. The extracellular matrix gel (called extracellular matrix gel or matrigel) was coated on the surface of the stamp, and then the matrigel patterned microarray was laid on the bottom of the non-adherent cell culture dish using the PDMS micropattern printing techno...

Embodiment 1

[0045] Example 1. The method for culturing lung tumor organoids of the present invention

[0046] The method for culturing lung tumor organoids of the present invention comprises the following steps:

[0047] (1) Preparation of porcine lung decellularized extracellular matrix glue: After taking fresh porcine lung, the pulmonary artery was cannulated, and the blood was removed by washing with PBS for 10 minutes. Lungs were then frozen and thawed, perfused with 1% Triton X-100 for 3 hours, followed by 1% sodium laureth sulfate (SLES) for 6 hours, then 1% Triton X-100 for 3 hours, and finally with Decellularized lungs were equilibrated by PBS perfusion for 2 hours to obtain decellularized lung scaffolds (lung extracellular matrix). During perfusion, the entire flow rate was 100 mL / min. Decellularized lung scaffolds (DLSs) were then cut into 1 × 1 × 1 cm cubes for lyophilization. Further crushed by a ball mill to obtain lung decellularized extracellular matrix powder. The abov...

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Abstract

The invention provides a method for culturing lung tumor organoids, and belongs to the field of organoid culture. The invention firstly provides a lung tumor organoid culture container which is a cell culture container with a lung extracellular matrix imprinting block array on the bottom surface in the lung tumor organoid culture container; the area, not covered with the lung extracellular matrix, of the bottom surface is sealed by a sealing agent; the lung extracellular matrix imprinting block array is composed of circular array points with the diameter of 50-200 [mu] m, and the distance between every two circular array points is 25-300 [mu] m. The invention further provides a culture method of the lung tumor organoid, the culture time is shortened, the culture mode is simplified, the particle size and arrangement of the lung tumor organoid are effectively controlled, and the repeatability and stability of drug screening are improved. The cultured lung tumor organoid is used for drug screening, higher efficiency and safety are achieved, meanwhile, personalized treatment can be conducted, and clinical medication is guided.

Description

technical field [0001] The invention belongs to the field of organoid culture, in particular to a method for culturing lung tumor organoids. Background technique [0002] Lung cancer is the malignant tumor with the highest morbidity and mortality in the world. According to statistics from the World Agency for Research on Cancer (IARC), in 2012, there were about 1.825 million new cases of lung cancer worldwide and about 1.59 million deaths. In my country, there were about 653,000 new cases of lung cancer and about 597,000 deaths. The most important malignant tumor that endangers the life and health of residents. With the further acceleration of industrialization and urbanization in my country, the air pollution and smoking rate remain high, and the harm of lung cancer will continue to increase. With the continuous development of science and technology, the precise treatment of tumors has entered the era of individualization: one is to use high-throughput gene sequencing to d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071C12M3/04C12M1/22C12M1/24C12Q1/02
CPCC12N5/0693C12N5/0688C12M23/10C12M23/08G01N33/5011C12N2503/02G01N2500/10C12N2533/90
Inventor 包骥步宏朱星龙李奕
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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