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Human skin epidermal cell culture medium and application thereof

A technology of epidermal cells and medium, which is applied in the field of biomedicine, can solve the problems of expensive addition of factors, time cannot meet the treatment of burns and scalds, slow expansion of epidermal cells, etc., and achieve the effect of shortening the time

Inactive Publication Date: 2016-09-21
JINAN PANSHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional epidermal medium not only needs expensive added factors, but also the expansion speed of the cultured epidermal cells is slow, which cannot meet the needs of burn and scald treatment in time. This factor seriously limits the application of tissue engineered epidermis in the treatment of acute skin wounds. Clinical application

Method used

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  • Human skin epidermal cell culture medium and application thereof
  • Human skin epidermal cell culture medium and application thereof
  • Human skin epidermal cell culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0041] Example 1: Comparison of the morphology and cell quantity of the original epidermis cultured by the human skin epidermal cell culture medium of the present invention and the traditional serum-containing epidermis medium

[0042] Materials and methods:

[0043] Tissue source: discarded neonatal foreskin during hospital operation

[0044] Cell culture medium: traditional serum-containing epidermal medium, human skin epidermal medium of the present invention

[0045] a) The collected fresh tissue samples were disinfected with alcohol for three minutes, and then treated twice with PBS containing 2 times double antibody, each time for 3 minutes. The tissues were soaked in F12 medium containing 1 times double antibody and stored at 4°C for later use. Tissue storage time is not higher than 72 hours.

[0046] b) Take out the tissue in the soaking liquid, drain the excess liquid, weigh it and make a record, cut the tissue into long strips, spread it evenly in a 100mm petri di...

example 2

[0055] Example two: adopt human skin epidermal cell culture medium of the present invention and the comparison of the original epidermal cell morphology and cell quantity of traditional serum-free keratin medium (K-sfm) culture

[0056] Materials and methods:

[0057] Tissue source: discarded fetal scalp during hospital operation

[0058] Cell culture medium: K-sfm medium, human skin epidermis medium of the present invention

[0059] a) The collected fresh tissue samples were disinfected with alcohol for three minutes, and then treated twice with PBS containing 2 times double antibody, each time for 3 minutes. The tissues were soaked in F12 medium containing 1 times double antibody and stored at 4°C for later use. Tissue storage time is not higher than 72 hours.

[0060] b) Take out the tissue in the soaking liquid, drain the excess liquid, weigh it and make a record, cut the tissue into long strips, spread it evenly in a 100mm petri dish, and add 2.5mg / ml Neutral protease...

example 3

[0069] Example three: adopt human skin epidermis culture medium of the present invention and the comparison of the passage newborn's foreskin epidermal cell morphology and cell quantity that traditional serum-free keratin medium (K-sfm) cultivates

[0070] Materials and methods:

[0071] Cells: subcultured neonatal foreskin epidermal cells

[0072] Medium: K-sfm medium, human skin epidermis medium of the present invention

[0073] a) Remove the old culture medium in the culture bottle.

[0074] b) Wash each bottle once with 5-10ml PBS to remove residual serum.

[0075] c) Add 3 ml of 0.05% trypsin containing EDTA to T75flask and digest in a 37 degree incubator for 10 min.

[0076] d) Observe under the microscope, if the cells are all detached, add 10ml of DMEM containing 10% FBS to each bottle for neutralization (can be gently blown several times), and collect all the cells into a 50ml centrifuge tube.

[0077] e) Centrifuge at 1000 rpm for 5 minutes, and discard the super...

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Abstract

The invention provides a human skin epidermal cell culture medium and application thereof. The culture medium is prepared by the following steps: mixing a K-SFM (keratinocyte serum-free medium), a DMEM (dulbecco's modified eagle medium) and a F12 culture medium according to the ratio of 2:1:1, and adding a BPE (bovine pituitary extract), an EGF (epidermal growth factor), an SCGF (stem cell growth factor), an FGF (fibroblast growth factor), a TGF-beta (transforming growth factor-beta), a VEGF (vascular endothelial growth factor), CaCl2, glutamine and a double-antibody. The culture medium is free of serum, and can be used for human skin epidermal cell primary culture and subculture. The human skin epidermal cell culture medium can obtain abundant epidermal cells by in-vitro culture by using only a small amount of patient skin tissues. Compared with the traditional epidermal cell culture medium, the human skin epidermal cell culture medium provided by the invention greatly shortens the time required by amplifying the same cell count, and satisfies the demands for clinical therapy.

Description

technical field [0001] The invention relates to a human skin epidermal cell culture medium and application thereof, belonging to the field of biomedicine. Background technique [0002] The skin is the largest organ in the human body. It undertakes functions such as protecting the body, perspiration, and feeling cold, heat, and pressure, and protects various tissues and organs in the body from physical, mechanical, chemical, and pathogenic microorganisms. Genetic diseases, burns, chronic skin trauma (such as diabetes), vitiligo, albinism and many other reasons can cause damage to the skin, affect the appearance of the skin, leave scars on the skin and even lose its physiological function. When the skin is seriously damaged, such as deep and large-scale burns, the skin can no longer recover by itself, and will lose its resistance to bacteria, causing a large number of bacteria to invade the human body through the skin, which can easily cause systemic infection and cause death....

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0625C12N2500/14C12N2500/30C12N2500/46C12N2500/84C12N2500/90C12N2501/11C12N2501/113C12N2501/125C12N2501/15C12N2501/165
Inventor 邢志青吴训伟
Owner JINAN PANSHENG BIOTECH
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