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Multifunctional serum-free cell culture medium and application thereof

A culture medium and multi-functional technology, applied in the field of biomedicine, can solve the problem of single use of cell culture medium, achieve good growth status, avoid impact, and fast adherence

Inactive Publication Date: 2016-08-17
JINAN PANSHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In tissue-engineered skin, the existing cell culture medium is single-purpose and has great limitations
At present, there is no medium that can be used not only for the rapid expansion of dermal fibroblasts in vitro, but also for the culture of primary epidermal cells

Method used

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  • Multifunctional serum-free cell culture medium and application thereof
  • Multifunctional serum-free cell culture medium and application thereof
  • Multifunctional serum-free cell culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, a kind of multifunctional serum-free cell culture medium, required reagent and consumption are as follows:

[0051]

[0052] In the ultra-clean workbench, mix DMEM high-glucose medium and F12 nutrient medium, add appropriate amount of B27 supplement, PC-1 supplement, recombinant human epidermal growth factor and penicillin, and mix well. The prepared medium can be stored stably at 4°C for 3-4 months.

Embodiment 2

[0053] Example 2, the multifunctional serum-free cell culture medium of the present invention is used for recovery and cultivation of frozen primary epithelial cells

[0054] (1) Pretreatment of the culture bottle with Coating Matrix: Add 0.5-3ml of Coating Matrix (Gibco, R011k) into the culture bottle and spread evenly, let it stand at room temperature for 10 minutes, and then discard the excess liquid in the bottle.

[0055] (2) Remove adult and embryo primary epithelial cells from a liquid nitrogen tank, and dissolve them by shaking gently in a 37°C water bath.

[0056] (3) The lysed cells were added to about 8 ml of DMEM containing 10% FBS, and centrifuged at 1000 rpm for 5 min.

[0057] (4) Discard the supernatant, resuspend the adult and embryonic cell pellets in 10ml of F12 medium containing double antibodies and wash the cells once, and centrifuge at 1000rpm for 5min.

[0058] (5) Discard the supernatant, add 10ml of the multifunctional serum-free cell culture medium ...

Embodiment 3

[0062] Embodiment 3, the multifunctional serum-free cell culture medium of the present invention is used for the cultivation of the primary epithelial cell of separation

[0063] (1) Pretreatment of the culture bottle with Coating Matrix: Add 0.5-3ml of the prepared Coating Matrix (Gibco, R011k) into the culture bottle and spread evenly, let it stand at room temperature for 10 minutes, and then discard the excess liquid in the bottle.

[0064] (2) The isolated adult and embryonic cells were resuspended and washed once in 10ml of F12 medium containing double antibody, and centrifuged at 1000rpm for 5min.

[0065] (3) Add 10ml of the multifunctional serum-free cell culture medium of the present invention to resuspend the adult cells and embryonic cells respectively, and add 0.5-50ul of 10uM Rock inhibitor to the culture medium, and inoculate them in T75 culture flasks. At this time, the cells were recorded as P0 generation.

[0066] (4) Place the culture bottle in an incubator ...

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Abstract

The invention provides a multifunctional serum-free cell culture medium and application thereof. A method for preparing the multifunctional serum-free cell culture medium includes mixing DMEM (dulbecco modified eagle medium) high-glucose culture media and F12 nutrient culture media with one another; adding B27 additives, PC-1 additives, gene recombination human epidermal growth factors and myllicin double-antibodies into the DMEM high-glucose culture media and the F12 nutrient culture media. The multifunctional serum-free cell culture medium and the application have the advantages that the multifunctional serum-free cell culture medium contains simple components, and repeated passage requirements can be met; more importantly, the multifunctional serum-free culture medium not only can be used for quickly amplifying and cultivating primary hypodermal fibroblasts and passage hypodermal fibroblasts in an in-vitro manner, but also can be used for cultivating primary epidermal cells.

Description

technical field [0001] The invention relates to a multifunctional serum-free cell culture medium and its application, 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 deat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N2500/90C12N2501/11
Inventor 邢志青吴训伟
Owner JINAN PANSHENG BIOTECH
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