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Corneal cell senescence model as well as establishment method and application thereof

A technology of cell aging and corneal epithelial cells, which is applied in the field of biomedicine, can solve the problems of difficult modeling success and long stimulation cycle, and achieve the effect of short time-consuming, simple equipment and high reproducibility of results

Pending Publication Date: 2021-12-10
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods either have a long stimulation period, are difficult to model successfully, or require the use of toxic reagents

Method used

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  • Corneal cell senescence model as well as establishment method and application thereof
  • Corneal cell senescence model as well as establishment method and application thereof
  • Corneal cell senescence model as well as establishment method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] 1. Test materials and equipment

[0124] Experimental materials: primary human corneal epithelial cells (HCEC, Qingqi Biotechnology Development Co., Ltd.), DMEM high-glucose medium (without sodium pyruvate, Gibco), fetal bovine serum (Gibco), trypsin (KGI Biotech) .

[0125] Main equipment: Inverted microscope (OLYMPUS, CKX41), ultra-clean workbench (Donglian Hal, SCB-1520), carbon dioxide cell incubator (SANYO), microplate reader (BIO-RAD, iMark), UV-8 ultraviolet light Light box (Beijing Electric Light Source Research Institute), ST-513 ultraviolet measuring instrument (Taiwan Xianchi), flow cytometer (BD, Accuri C6 PLUS), blue light irradiation lamp, blue light irradiation meter (Beijing Normal University Photoelectric Instrument Factory ).

[0126] 2. Test method

[0127] 2.1 Cell recovery

[0128] Take the purchased primary human corneal epithelial cells out of the dry ice, immediately place them in a 40°C water bath, melt them as soon as possible, and add 4 mL...

Embodiment 2

[0156] All experimental materials and test methods of this embodiment are consistent with those of Example 1, the only difference being that in the cell aging step, the cell culture plate was heated at 2.0J / cm 2 UVA and 50mJ / cm 2 After UVB irradiation, add 1000lux blue light for 1 hour.

Embodiment 3

[0158] All experimental materials and test methods of embodiment 3 are consistent with embodiment 1, the only difference is that in the cell aging step, the cell culture plate is heated at 7.2J / cm 2 UVA and 126mJ / cm 2 After UVB irradiation, add 2000lux blue light for 1 hour.

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Abstract

The invention provides a corneal cell senescence model. The corneal cell senescence model is characterized by being constructed by the following steps that a cell culture step is conducted: corneal cells are cultured by using a cell culture solution; a cell aging step is conducted: the cell culture solution after the cell culture step is discarded, a buffer solution is added, and the corneal cells are exposed under the conditions of UVA, UVB and blue light; and a re-culture step is conducted: the buffer solution is discarded, a serum-free culture solution is added, and the corneal cells are re-cultured to obtain the corneal cell senescence model. The invention also provides an establishment method of the corneal cell senescence model and a method for evaluating the anti-aging effect of a medicine on the corneal cells in vitro. According to the corneal cell senescence model as well as the establishment method and application thereof, the corneal cell senescence model is simple in equipment, easy to operate, short in consumed time and free of toxic and harmful reagents, and the good effect of high result reproducibility can be achieved.

Description

technical field [0001] The application relates to the field of biomedicine, in particular to a corneal cell aging model, a method for establishing the corneal cell aging model, and a method for evaluating the anti-aging effect of drugs on corneal cells in vitro. Background technique [0002] Statistics from the Chinese Medical Association show that with the development of science and technology and the needs of life and work, people spend more and more time on smartphones, computers, and TVs. By 2020, the number of smartphone users is expected to increase by 53%. 60% of people spend at least 6 hours a day on digital devices, and 63.5% of people suffer from vision loss, cataract, blindness and other eye diseases of varying degrees due to blue light radiation. Most of the liquid crystal displays equipped with these video display terminals use light-emitting diodes (LEDs) as backlight sources, and the blue light content in LEDs is extremely high, causing great damage to the cor...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N13/00C12Q1/02
CPCC12N5/0621C12N5/0625C12N13/00G01N33/5044C12N2503/02C12N2500/90C12N2529/10G01N2500/10
Inventor 毛华张晓鸥谢文娟李芬郭学平
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD