In-vitro simulated organoid cultivation method for colon cancer cells

A technology of colon cancer cells and culture methods, which is applied in the field of in vitro tissue culture of colon cancer cells, can solve problems such as increased expression levels of brush border enzymes, achieve perfect cell function analysis, promote uniform cell attachment, and promote rapid differentiation effect

Inactive Publication Date: 2016-07-13
ZHEJIANG UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The study found that the hydrophilic modified material is helpful for the attached growth of cells, but the expression level of the brush border enzymes of the cells grown on the surface of the hydrophobic material is relatively increased

Method used

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  • In-vitro simulated organoid cultivation method for colon cancer cells
  • In-vitro simulated organoid cultivation method for colon cancer cells
  • In-vitro simulated organoid cultivation method for colon cancer cells

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Embodiment 1

[0053] Embodiment 1, an in vitro tissue-like culture method of colon cancer cells (the construction method of the Caco-2 cell hollow fiber reactor used for functional component research), the following steps are carried out successively:

[0054] 1), Sterilization of hollow fiber:

[0055] Cut the hollow fiber into small pieces with a length of 3 cm, soak in PBS buffer solution (pH=7.4), and sterilize in a steam autoclave (1.1 atmospheres, 121°C) for 20 minutes;

[0056] After the sterilization is completed, take out the hollow fiber in the ultra-clean bench and place it on a plate to air dry, usually overnight;

[0057] The above-mentioned hollow fibers are PES hollow fiber membranes or PVDF hollow fiber membranes;

[0058] The PES (polyethersulfone) hollow fiber membrane has a pore size of 0.1 μm and a pure water flux of 1000 L / m at 0.1 MPa 2 h, inner / outer diameter 0.8 / 1.4mm;

[0059] The PVDF (polyvinylidene fluoride) hollow fiber membrane has a pore size of 0.2 μm and ...

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Abstract

The invention discloses an in-vitro simulated organoid cultivation method for colon cancer cells.The in-vitro simulated organoid cultivation method sequentially includes steps of sterilizing hollow fibers; preliminarily laying collagen on the hollow fibers; injecting Caco-2 cell suspension into inner cavities of the hollow fibers, arranging the hollow fibers in incubators at the temperatures of 37+/-0.2 DEG C, and driving the hollow fibers by driving devices to axially rotate at the rotational speeds of 90 degrees/hour for the rotational time of 4 h so as to uniformly attach cells; arranging the hollow fibers with the cultivated cells in cultivation plates, adding 2-3 mL of cultivation media into each hole in each cultivation plate, arranging the cultivation plates in the incubators at the temperatures of 37+/-0.2 DEG C and cultivating the cells; changing the cultivation media once every 1-2 days.The in-vitro simulated organoid cultivation method has the advantages that small intestinal cell hollow fiber membrane reactors capable of simulating micro-morphology of human tissues can be constructed by the in-vitro simulated organoid cultivation method, and the in-vitro simulated organoid cultivation method can be applied to research on functional component transport and absorption behavior.

Description

technical field [0001] The invention relates to an in vitro culture method of animal cells, in particular to an in vitro histochemical culture method of colon cancer cells; and to a construction method of a Caco-2 cell hollow fiber reactor for functional component research. Background technique [0002] Caco-2 cells are human colon adenocarcinoma cells with good homology and strong vitality. After about 3 weeks of culture, they can form a continuous cell monolayer and differentiate into nearly mature intestinal epithelial cells in terms of morphology and function. status [1] , and correspondingly express the role of small intestinal hydrolytic enzymes (such as maltase, lactase, aminopeptidase, etc.). [0003] As attachment-dependent cells, the monolayer adherent culture model is the most common culture model for Caco-2 cells. To improve polar cell culture, the researchers cultured Caco-2 cells on insert-type nested culture plates covered with permeable microporous membrane...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2531/00C12N2533/54
Inventor 郑晓冬徐冬冬
Owner ZHEJIANG UNIV
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