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Intestinal epithelium monolayer culture and characterization method based on mouse intestinal stem cells

A culture method and technology of intestinal epithelium, applied in the field of intestinal epithelial monolayer culture and characterization, can solve the problem of inability to reproduce cell-cell interaction, etc., achieve the effect of optimizing the identification method in vitro and clarifying the reliability

Inactive Publication Date: 2019-04-26
ZHEJIANG GONGSHANG UNIVERSITY
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Problems solved by technology

In the past, the in vitro cell models used for the above functional studies were composed of tumor cell lines composed of single cells. Although these genetically modified intestinal epithelial cell lines have functions of intestinal absorptive cells or secretory cells, the single cell composition is only Enables analysis of relatively simple functions that cannot reproduce cell-cell interactions in the real gut

Method used

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  • Intestinal epithelium monolayer culture and characterization method based on mouse intestinal stem cells
  • Intestinal epithelium monolayer culture and characterization method based on mouse intestinal stem cells

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Embodiment 1

[0027] Step 1. Dilute the composite viscose solution at 1:20 with pre-cooled calcium- and magnesium-ion-free phosphate buffer solution on ice, and then add 150 microliters per well to 48-well culture plate 1. The solution should completely cover the bottom of the well plate. , and then placed in a 37°C carbon dioxide incubator for 2 hours;

[0028] Step 2: Kill 10-week-old C57BL / 6 mice by cervical dislocation; dissect and cut open the abdominal cavity of the mice, pull out the small intestine from the abdominal cavity, take jejunum with a length of 10-20 cm, and put it into pre-cooled calcium- and magnesium-free In ionic phosphate buffer solution, ophthalmic forceps cut the jejunum longitudinally along the longitudinal axis, wash with pre-cooled calcium and magnesium ion-free phosphate buffer solution to remove the chyme in the intestine; Gently scrape off the villi of the small intestine, and wash the intestinal segment once with pre-cooled calcium- and magnesium-ion-free pho...

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Abstract

The invention discloses an intestinal epithelium monolayer culture and characterization method based on mouse intestinal stem cells. The method comprises the following steps: (1) monolayer culture ofmouse intestinal epithelium; (2) characterization of monolayer culture of the intestinal epithelium. Compared with the prior art, the method is based on small intestinal crypts containing stem cells,optimizes the monolayer in-vitro culture system and culture medium of the intestinal epithelium, and includes a complete culture and characterization method.

Description

technical field [0001] The invention relates to a method for culturing and characterizing intestinal epithelial monolayers based on mouse intestinal stem cells. Background technique [0002] As the largest nutrient recognition and absorption organ in the human body, the intestinal tract has an important guiding role in the prevention and treatment of metabolic disorders such as obesity and diabetes regarding the recognition, perception, transport and hormone release of food at the intestinal epithelial level. In the past, the in vitro cell models used for the above functional studies were composed of tumor cell lines composed of single cells. Although these genetically modified intestinal epithelial cell lines have the function of intestinal absorptive cells or secretory cells, the single cell composition is only Can be used for analysis of relatively simple functions, unable to reproduce cell-cell interactions in the real intestine. Fortunately, the discovery of intestinal...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071G01N33/543G01N33/531G01N21/64
CPCC12N5/0679G01N21/6458G01N33/531G01N33/54306
Inventor 秦玉梅韩剑众王稣嫱
Owner ZHEJIANG GONGSHANG UNIVERSITY
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