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Dry cell model and establishment method and application thereof

A technology for cell model and method establishment, which can be applied to artificial cell constructs, animal cells, vertebrate cells, etc., and can solve the problems of difficult oven temperature, unstable ultra-clean typhoon speed, and great influence on the stability of experimental results.

Active Publication Date: 2019-09-24
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to maintain the temperature accurately in the oven. Generally, the minimum temperature of the oven is the ambient temperature plus 10°C. In the hot summer, the oven temperature is generally 40-45°C. Drying at this temperature will cause a certain degree of thermal damage to the cells.
The wind drying method generally uses the wind of the ultra-clean bench to dry and damage the cells in the ultra-clean bench. The disadvantage of this method is that the ultra-clean bench with vertical airflow is required, and the wind speed of the ultra-clean bench is unstable and uniform. Poor, great impact on the stability of the experimental results

Method used

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  • Dry cell model and establishment method and application thereof
  • Dry cell model and establishment method and application thereof
  • Dry cell model and establishment method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0139] Experimental example 1 Desiccator method prepares the dry cell model of human skin fibroblast

[0140] 1.1 Test material

[0141] Experimental materials: human skin fibroblasts, RPMI-1640 medium, fetal bovine serum (FBS), trypsin.

[0142] Main equipment: glass desiccator (diameter 300mm), temperature and hygrometer, inverted microscope, ultra-clean workbench, carbon dioxide incubator.

[0143] 1.2 Experimental method and process

[0144] 1.2.1 Preparation before experiment

[0145] Prepare a glass desiccator and add 500 g of silica gel to dry the particles (4 hours at 120° C. in an oven). Seal it for 1 hour, measure the relative humidity in the desiccator to be 10.0%, the temperature is 25° C., and irradiate for 1 hour under the ultraviolet lamp.

[0146] 1.2.2 Planking

[0147] Take 5×10 4 Cells / mL The recovered human skin fibroblasts of the third generation were inoculated on a 96-well plate, 125 μL per well, and 5 replicate wells were set for each sample. At ...

experiment example 2

[0159] Experimental Example 2 Desiccator Method Preparation of Dry Cell Model of Human Skin Fibroblasts

[0160] 2.1 The specific parameters are the same as in Experimental Example 1, where the drying time in 1.2.3 is replaced with 30 minutes.

[0161] 2.2 Results and analysis

[0162] The experiment was repeated three times, and the cell viability is shown in Table 2 below.

[0163] Table 2 Cell viability (%)

[0164]

[0165] Note: ** means p<0.01 compared with the non-dried treatment group

[0166] Conclusion: It is feasible to use a desiccator to dry and damage cells, and the error of three parallel experiments is within 5.0%. The drying damage effect of Experimental Example 2 is more significant.

experiment example 3

[0167] Experimental example 3 Desiccator method prepares human skin keratinocyte drying model

[0168] 3.1 Experimental materials

[0169] Experimental materials: human skin keratinocytes (HaCaT), DMEM high-glucose medium, fetal bovine serum, trypsin.

[0170] Main equipment: glass desiccator (300mm in diameter), inverted microscope, ultra-clean bench, carbon dioxide incubator, temperature and humidity meter

[0171] 3.2 Experimental method and process

[0172] 3.2.1 Preparation before experiment

[0173] Prepare a glass desiccator and add 500 g of silica gel to dry the particles (4 hours at 120° C. in an oven). Seal it for 1 hour, measure the relative humidity in the desiccator to be 9.0%, the temperature is 26°C, and irradiate for 1 hour under the ultraviolet lamp.

[0174] 3.2.2 Planking

[0175] Take 5×10 5 Seed the human skin keratinocytes of the third passage after resuscitation in a 96-well plate at a density of cells / mL, 100 μL per well, 37 ° C, 5% CO 2 Conditio...

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Abstract

The invention provides a dry cell model establishment method. The method includes the following steps of cell culture, cell drying and re-culture; in the cell culture step, cells are cultured on a cell culture plate under the conditions of the controlled temperature and humidity; in the cell drying step, the cells are exposed in a cell culture solution obtained after cell culture under the conditions that the wind speed is less than 0.1m / s, the relative humidity is 10.0% or less and the temperature is at 25+ / -5 DEG C; in the re-culture step, the cells are re-cultured under the conditions of the controlled temperature and humidity, and the activity of the dried cells is reduced to be 45-70%, wherein the cell activity is measured by a WST-1 method, the absorbance value of an un-dried set obtained after incubation under given conditions is defined as 100%, the absorbance value of a dried set obtained after the same incubation under given conditions is measured, and the detection result of the absorbance value of the un-dried set to the absorbance value of the dried set is obtained and represented in percentage. The method can be used for evaluating the dry protection and / or dry repair performance of cosmetics and their raw materials.

Description

technical field [0001] The invention relates to a method for evaluating the moisturizing properties of cosmetics and their raw materials, in particular to a method for evaluating the moisturizing properties of cosmetics and their raw materials and their protective and repairing effects on dry damage at the cell level. Background technique [0002] Moisturizing is the basic appeal of skin care, and moisturizing effect is the most basic function of cosmetics. Therefore, cosmetics claiming moisturizing occupy the highest proportion in the entire cosmetic category. Researchers are also constantly introducing new mechanisms of action and new evaluation methods for moisturizing efficacy. The "Guidelines for the Evaluation of Cosmetic Moisturizing Efficacy" is the first domestic industry standard on the evaluation of cosmetic efficacy, which plays a role in regulating the order of the cosmetics market and improving the quality of cosmetics. This method is to measure the water cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N5/0629
Inventor 毛华张晓鸥谢文娟王玉玲陈雯雯范馨仪石艳丽郭学平
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD