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Cell culture medium, cell culture kit and cell culture method

A cell culture and culture medium technology, applied in the field of cell culture medium, can solve the problems of fast aging and death, death and other problems

Active Publication Date: 2020-05-01
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]However, normal human sebocytes can only be cultured for 3-6 generations at the primary stage, and they will die in large numbers when they are cultured for 7-9 generations, and the in vitro expansion and passage should not exceed 10 Aging and even death will occur in the first generation, especially in the culture system containing serum, the aging and death speed is faster, this phenomenon cannot meet the needs of large-scale and long-term sebocyte experiments

Method used

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  • Cell culture medium, cell culture kit and cell culture method
  • Cell culture medium, cell culture kit and cell culture method
  • Cell culture medium, cell culture kit and cell culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] This example provides a sebocyte culture medium, which contains DMEM / Ham'F-12 (1 / 1), 0.2 mg / mL casein, 1.2 μg / mL transferrin, 12 μg / mL mL of bovine insulin, 1 μg / mL of linoleic acid, 400nmol / L of testosterone, 0.12mmol / L of vitamin A, 0.04mmol / L of L-ascorbic acid, 0.13mmol / L of polylysine, 0.4nmol / L of putrescine and 25 μmol / L of 2-mercaptoethanol.

[0097] The preparation method of the sebocyte culture medium of the present embodiment is as follows:

[0098] In a sterile clean bench, add 12 μg / mL bovine insulin, 0.2 mg / mL casein, 1.2 μg / mL trans Ferritin, 1μg / mL linoleic acid, 400nmol / L testosterone, 0.12mmol / L vitamin A, 0.04mmol / L L-ascorbic acid, 0.13mmol / L polylysine, 0.4nmol / L putrid Amine and 25 μmol / L 2-mercaptoethanol, mixed by pipetting, and sterilized by positive pressure filtration with a microporous filter membrane with a pore size of 0.22 μm.

Embodiment 2

[0100] This example provides a sebocyte culture medium, which contains DMEM / Ham'F-12 (1 / 1), 5 mg / mL casein, 5 μg / mL transferrin, 30 μg / mL Bovine insulin, 5μg / mL linoleic acid, 800nmol / L testosterone, 0.28mmol / L vitamin A, 0.28mmol / L L-ascorbic acid, 0.26mmol / L polylysine, 2.5nmol / L putrid amine and 120 μmol / L 2-mercaptoethanol.

[0101] The preparation method of the sebocyte culture medium of the present embodiment is as follows:

[0102] In a sterile clean bench, add 30 μg / mL bovine insulin, 5 mg / mL casein, and 5 μg / mL transferrin to the culture medium with a volume ratio of DMEM:Ham'F-12 of 1:1 , 5μg / mL of linoleic acid, 800nmol / L of testosterone, 0.28mmol / L of vitamin A, 0.28mmol / L of L-ascorbic acid, 0.26mmol / L of polylysine, 2.5nmol / L of putrescine and 120 μmol / L 2-mercaptoethanol, pipette and mix well, and use a microporous filter membrane with a pore size of 0.22 μm to filter and sterilize under positive pressure.

Embodiment 3

[0104] This example provides a sebocyte culture medium, which contains DMEM / Ham'F-12 (1 / 1), 0.5 mg / mL casein, 1.5 μg / mL transferrin, 15 μg / mL mL of bovine insulin, 5μg / mL of linoleic acid, 400nmol / L of testosterone, 0.15mmol / L of vitamin A, 0.05mmol / L of L-ascorbic acid, 0.26mmol / L of polylysine, 1nmol / L of Putrescine and 25 μmol / L 2-mercaptoethanol.

[0105] The preparation method of the sebocyte culture medium of the present embodiment is as follows:

[0106] In a sterile clean bench, add 15 μg / mL bovine insulin, 0.5 mg / mL casein, 1.5 μg / mL trans Ferritin, 5 μg / mL linoleic acid, 400 nmol / L testosterone, 0.15 mmol / L vitamin A, 0.05 mmol / L L-ascorbic acid, 0.26 mmol / L polylysine, 1 nmol / L putrescine and 25 μmol / L 2-mercaptoethanol, mixed by pipetting, and sterilized by positive pressure filtration with a microporous filter membrane with a pore size of 0.22 μm.

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Abstract

The invention relates to a cell culture medium. The cell culture medium contains a basal culture medium, a serum substitute, linoleic acid, testosterone, vitamin A, L-ascorbic acid, polylysine, putrescine, and 2-mercaptoethanol. The cell culture medium of the invention can efficiently culture sebaceous gland cells in vitro, so that a cell proliferation rate, a survival rate after resuscitation, and an expression amount of lipid synthesis related proteins of different generations of the sebaceous gland cells are significantly increased, and thereby the assistance for extensively researching sebaceous gland physiological and endocrine functions, exploring pathogenesis of sebaceous gland related diseases, and developing drugs is provided.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a cell culture medium, a cell culture kit and a cell culture method. Background technique [0002] Sebaceous glands are branched polyglands, which exist in all parts of the human skin except the palms, soles, and dorsums of the feet, especially concentrated on the scalp, forehead, and face. There may be 400-900 glands per square centimeter area. Sebaceous gland cells exist in the sebaceous glands, which play a role in releasing sebum. Sebocytes differentiate and divide to become fully mature cells that produce and secrete an oily, waxy substance (sebum) whose main components include triglycerides and fatty acid breakdown products, wax esters, squalene, cholesteryl esters, and cholesterol, Protects and moisturizes skin and hair. Abnormal secretion of sebum is the cause of skin diseases such as oily skin, dry skin, acne, seborrhea, seborrheic dermatitis, and xeroderma. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2500/46C12N2500/44C12N2500/32C12N2500/38C12N2500/30C12N2500/24C12N2501/33C12N2501/998
Inventor 杨文娟卢永波张勇杰李潇
Owner GUANGDONG BOXI BIO TECH CO LTD
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