Method for in-vitro induced decidualization of mouse uterus primary stroma cells

A stromal cell and uterine technology, applied to artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of high mortality, low success rate, and difficulty in starting decidualization, so as to ensure the survival rate and success rate High, promote the effect of starting

Active Publication Date: 2019-05-21
NORTHWEST NORMAL UNIVERSITY
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  • Summary
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for inducing decidualization of mouse uterine primary stromal cells in vitro, to solve the high mortality rate of mouse uterine primary stromal cells in the prior art, difficulty in starting decidualization, and success low rate of defects

Method used

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Embodiment 1

[0016] A method for inducing decidualization of mouse uterine primary stromal cells in vitro, comprising the following steps:

[0017] (1) After the mice on the 4th day of normal pregnancy were killed, they were sterilized in 75% alcohol for 2 minutes, and the abdominal cavity was opened under aseptic conditions. Make a small cut at the cervix, cut open the uterine cavity on both sides longitudinally with scissors, and collect the uterus. Try to remove all the mesentery and fat on one side of the uterus and put them in a 15ml centrifuge tube filled with PBS buffer. Shake vigorously to remove For blood and other tissues, the uterine tissue of the mouse was obtained, and trypsin solution was added to it, and then digested in a 4°C refrigerator and a 37°C cell culture incubator for 40 minutes, and then the digestion reaction was terminated with 10% PBS solution , remove the epithelial cells, and obtain the tissue block, wherein, the trypsin solution is made by dissolving trypsin ...

Embodiment 2

[0021] A method for inducing decidualization of mouse uterine primary stromal cells in vitro, comprising the following steps:

[0022] (1) After killing the mice on the 4th day of normal pregnancy, they were sterilized in 75% alcohol for 2 minutes, and the abdominal cavity was opened under aseptic conditions. Make a small cut at the cervix, use scissors to cut the uterine cavity longitudinally on both sides and collect the uterus, try to remove all the mesentery and fat on one side of the uterus and put it in a 15ml centrifuge tube filled with PBS buffer, shake it vigorously to Remove blood and other tissues to obtain mouse uterine tissue, add trypsin solution to it, put it in a refrigerator at 4°C and a cell culture incubator at 37°C for 40 minutes, and then use 10% PBS solution to terminate the digestion reaction Reaction, removing epithelial cells, and obtaining tissue pieces, wherein, the trypsin solution is made by dissolving trypsin and EDTA in a calcium-magnesium-free b...

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Abstract

The invention discloses a method for in-vitro induced decidualization of mouse uterus primary stroma cells. The method includes the following operation steps of firstly, taking a tissue block of uterus tissue of a mouse in the fourth day after the mouse normally gives birth; secondly, digesting the tissue block to obtain a single-cell suspension of the mouse uterus primary stroma cells; thirdly, culturing the single-cell suspension until the cell density reaches 75-85% for combination, exchanging fluid, removing parenchyma cells, and adding an induction medium for inducing the decidualizationof the mouse uterus primary stroma cells. The method is simple in operation, the mouse uterus primary stroma cells separated out can have a high survival rate, meanwhile the decidualization startup success rate is high, and a foundation is laid for the research of researches on the decidualization mechanism.

Description

technical field [0001] The invention relates to the technical field of cell induction, and specifically relates to a method for inducing decidualization of mouse uterine primary stromal cells in vitro. Background technique [0002] With the development of industrialization and the great change of human life style, the proportion of infertility patients in the world has been increasing year by year in recent years. At present, although artificial assisted pregnancy technology has been greatly developed, even in the case of high-quality embryos, there are still problems of low implantation rate of embryo transfer and low success rate of subsequent pregnancy, which is mainly due to Membrane growth and development disorders. The success of embryo implantation, on the one hand, depends on the early embryo development to form a blastocyst with implantation ability, and on the other hand, it depends on the endometrium entering into a receptive state, that is, the "window period" o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 张晋平谭毅丁兰丁艳平孔维宝王俊龙罗文萍傅龙飞马君义
Owner NORTHWEST NORMAL UNIVERSITY
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