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In-vitro organ culture method for embryo kidneys

A technology of organ culture and kidney, which is applied in the biological field, can solve the problems that the culture method requires special materials and occupies a large space, and achieves simple and easy pollution links, overcomes the large space occupation, and achieves good results in culture

Inactive Publication Date: 2012-03-28
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is mainly the defect that the existing culture method requires special materials and occupies a large space

Method used

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  • In-vitro organ culture method for embryo kidneys
  • In-vitro organ culture method for embryo kidneys
  • In-vitro organ culture method for embryo kidneys

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Using the method of the present invention to cultivate the kidneys of Balb / c mice at embryonic day 11.5.

[0054] 1. Prepare the carrier: use a glass dome with a diameter of 12mm as the carrier, and buy it from Haimen Huakai Experimental Glass Instrument Co., Ltd. First, prepare a 1% hydrochloric acid solution with distilled water, shake and clean the dome at room temperature for one hour. After cleaning with distilled water, put it into a 70% alcohol solution by volume, shake and clean overnight. Use a 10cm glass plate to hold the cleaned domes, drain the liquid, put it in a high-temperature steam sterilizer, and sterilize at 121°C for 20 minutes. After drying, put it on the ultra-clean workbench for later use.

[0055] 2. Assemble the culture device: use sterile tweezers to pick up the processed dome and place it in the culture well of a sterile 24-well culture plate. The sterile 24-well culture plate was 3524 tissue culture plate purchased from Costar (C...

Embodiment 2

[0067] Embodiment 2 Transfection of the embryonic kidney obtained by the culture method of the present invention

[0068] 1. Prepare the carrier sheet: a glass dome with a diameter of 12 mm is used as the carrier sheet, purchased from Haimen Huakai Experimental Glass Instrument Co., Ltd. First, prepare a 1% hydrochloric acid solution with distilled water, shake and clean the dome at room temperature for one hour. After cleaning with distilled water, put it into a 70% alcohol solution by volume, shake and clean overnight. Use a 10cm glass plate to hold the cleaned domes, drain the liquid, put it in a high-temperature steam sterilizer, and sterilize at 121°C for 20 minutes. After drying, put it on the ultra-clean workbench for later use.

[0069] 2. Assemble the culture device: Use sterile tweezers to pick up the treated round cover and place it in a sterile 24-well culture plate. The sterile 24-well culture plate is the 3524 tissue culture plate purchased from Costar Company...

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Abstract

The invention belongs to the biotechnology field, relates to an in-vitro organ culture method for embryo kidneys of mammals, and particularly relates to the isolated organ culture for the kidneys in embryonic periods of mice. The in-vitro organ culture method solves the main technical problems that a conventional culture method has the disadvantages of needing special materials and occupying large space. The in-vitro organ culture method for the embryo kidneys adopts the technical scheme that the method comprises the steps as follows: firstly, carrying tablets are put in cultivation holes formed on a porous culture plate and are matched with the cultivation hole in size, and a cultivation space is formed at the bottom part of each cultivation hole; secondly, obtained mammalian embryo kidneys or embryo kidney primordia are placed on the carrying tablets, culture mediums are added in, and the kidneys are led to be positioned at the junctions of the culture mediums and air; and thirdly, the kidneys are cultured in an incubator under the temperature of 37 DEG C, and then solution changing of the culture mediums is carried out every one day or every two days. By adopting the in-vitro organ culture method, large number of in vitro cultured kidney organs that are grown stably and are convenient for subsequent operation can be obtained at low cost, and the requirements of large-scale drug screening can be satisfied.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an in vitro organ culture method of mammalian embryonic kidney, in particular to an isolated organ culture method for mouse embryonic kidney. Background technique [0002] In vitro organ culture is an advanced form of tissue culture. It can reproduce the development process of organs and simulate the functional state of organs in different states and conditions. It is an important technology for understanding organ development process, researching disease pathogenic mechanism and drug screening. means. [0003] At present, there are currently kidney organ cultures in vitro: Cell contact and cell adhesion during tissue organization 1966May 15; 1 (3): 271-90, published in the International Journal of Cancer by Saxen L et al. , put in 2ml of culture medium, and float a polycarbonate filter membrane with a pore size of about 5um on the culture medium or use a stainless steel mesh as a sup...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
Inventor 周钦吕小岩陈铁林孙环魏于全
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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