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Cellular injury model preparation method based on ultraviolet irradiation

A cell damage and ultraviolet technology, applied in the field of cell experiments, can solve the problems of the large difference between the model and the human skin cell model and the low preparation success rate, and achieve the effect of improved survival rate, accurate experimental data and good proliferation.

Inactive Publication Date: 2020-01-14
FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for preparing a cell damage model based on ultraviolet irradiation, so as to solve the problem that the model prepared by the existing cell damage model preparation method proposed in the above-mentioned background technology is quite different from the human skin cell model, and the success rate of preparation is low. The problem

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for preparing a cell damage model based on ultraviolet radiation, the method comprising the following steps:

[0022] S1: Cut out the normal human skin tissue in the clinical operation, and disinfect it with povidone iodine. After the disinfection is completed, use 0.25% trypsin digestion solution to digest the skin tissue, disperse the tissue into single cells, and then put the cell solution in a suitable medium , place the culture medium in an incubator to allow these primary cells to grow and proliferate;

[0023] S2: After about 24 hours, the primary cells adhered to the wall and overgrown with the medium, then digested with 0.25% trypsin digestion solution, replaced with new medium and seeded on multiple six-well plates, the seeding density of the six-well plates About 5X10 4 / ml, each six-well plate after inoculation was placed in an incubator and cultured until it adhered to the wall, and then replaced with a new medium for further experiments. Multiple...

Embodiment 2

[0031] A method for preparing a cell damage model based on ultraviolet radiation, the method comprising the following steps:

[0032] S1: Cut out the normal human skin tissue in the clinical operation, and disinfect it with povidone iodine. After the disinfection is completed, use 0.25% trypsin digestion solution to digest the skin tissue, disperse the tissue into single cells, and then put the cell solution in a suitable medium , place the culture medium in an incubator to allow these primary cells to grow and proliferate;

[0033] S2: After about 24 hours, the primary cells adhered to the wall and overgrown with the medium, then digested with 0.25% trypsin digestion solution, replaced with new medium and seeded on multiple six-well plates, the seeding density of the six-well plates About 4X10 4 / ml, each six-well plate after inoculation was placed in an incubator and cultured until it adhered to the wall, and then replaced with a new medium for further experiments. Multiple...

Embodiment 3

[0041] A method for preparing a cell damage model based on ultraviolet radiation, the method comprising the following steps:

[0042]S1: Cut out the normal human skin tissue in the clinical operation, and disinfect it with povidone iodine. After the disinfection is completed, use 0.25% trypsin digestion solution to digest the skin tissue, disperse the tissue into single cells, and then put the cell solution in a suitable medium , place the culture medium in an incubator to allow these primary cells to grow and proliferate;

[0043] S2: After about 24 hours, the primary cells adhered to the wall and overgrown with the medium, then digested with 0.25% trypsin digestion solution, replaced with new medium and seeded on multiple six-well plates, the seeding density of the six-well plates About 6X10 4 / ml, each six-well plate after inoculation was placed in an incubator and cultured until it adhered to the wall, and then replaced with a new medium for further experiments. Multiple ...

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Abstract

The invention discloses a cellular injury model preparation method based on ultraviolet irradiation. The method comprises the following steps of: cutting a human body normal skin tissue in a clinicaloperation, and carrying out disinfection through iodophor; after the disinfection is finished, using 0.25% trypsin digestive juice to digest the skin tissue to disperse the tissue into single cells; then, placing cell sap in a proper culture medium, and placing the culture medium in an incubator for culture to enable primary cells to grow and proliferate; after about 24h, enabling the primary cells to grow all over the culture medium by attaching to a wall; then, carrying out digestion by 0.25% trypsin digestive juice; and replacing a new culture medium, and inoculating to a plurality of six-hole plates, wherein inoculation density on each six-hole plate is about 5*10<4> / ml. By use of the method, normal human body tissues are selected to successfully carry out culture in vitro, a high-purity human body normal cell is obtained, the prepared cellular injury model is more similar to a state obtained after a human skin is irradiated by ultraviolet rays, and obtained experiment data is moreaccurate.

Description

technical field [0001] The invention relates to the technical field of cell experiments, in particular to a method for preparing cell damage models based on ultraviolet radiation. Background technique [0002] Ultraviolet radiation is a low-energy electromagnetic radiation that has various biological effects, such as sterilization, inflammation, and carcinogenesis. Occupational and daily exposure to a certain amount of ultraviolet radiation can lead to disorders of the human neuroendocrine system and decline in immune function. The skin, like other human organs, ages over time, but it is different from other organs. An important environmental factor that affects skin aging is ultraviolet radiation. With the increase of atmospheric ozone holes, the aggravation of ambient light pollution and people's living standards People are paying more and more attention to the problems of erythema, wrinkles, pigmentation, and weakened skin elasticity caused by photodamage. Most of the exp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N13/00
CPCC12N5/0625C12N13/00
Inventor 徐丹张娟涂颖颜光前起珏崔婷婷
Owner FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV