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Serum-free culture medium for induced pluripotent stem cells and preparation method of serum-free culture medium

A serum-free medium, pluripotent stem cell technology, applied in artificially induced pluripotent cells, cell culture active agents, biochemical equipment and methods, etc. Instability and other problems, to achieve the effect of shortening the cell division cycle, maintaining proliferation activity, and maintaining a good growth state

Inactive Publication Date: 2021-05-28
齐国友
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Induced pluripotent stem cells produced by traditional culture conditions cannot be used as a suitable cell source because they are prone to human rejection after transplantation due to long-term exposure to a culture environment rich in animal products
At the same time, the traditional culture conditions are still very unstable and cannot meet the needs of experimental and clinical applications.

Method used

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  • Serum-free culture medium for induced pluripotent stem cells and preparation method of serum-free culture medium

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A serum-free medium for induced pluripotent stem cells, consisting of basal medium, adding the following components in the basal medium: insulin, diclofenazine, heparan sulfate glycoprotein, furadienes, D-menthone, Composed of kavalactone, IFN-γ, EGF, and zinc citrate; the dosage of each component in terms of final concentration is as follows: insulin 12 μg / mL, diclofenazine 3 μg / mL, heparan sulfate glycoprotein 18 μg / mL, difuran ene 3ng / mL, D-menthone 5ng / mL, kavalactone 12μg / mL, IFN-γ8ng / mL, EGF12ng / mL, zinc citrate 15μg / mL.

[0023] The preparation method of the above-mentioned serum-free medium for inducing pluripotent stem cells comprises the following steps:

[0024] (1) Add insulin, diclofenazine, heparan sulfate glycoprotein, IFN-γ, EGF to the basal medium and stir to mix evenly;

[0025] (2) Add furadienes, D-menthone, carbalactone, and zinc citrate to the mixture in step (1), continue to stir until evenly mixed, and then let it stand still, and use a 0.22 μm ...

Embodiment 2

[0027] A serum-free medium for induced pluripotent stem cells, consisting of basal medium, adding the following components in the basal medium: insulin, diclofenazine, heparan sulfate glycoprotein, furadienes, D-menthone, Composed of kavalactone, IFN-γ, EGF, and zinc citrate; the dosage of each component in terms of final concentration is as follows: insulin 10 μg / mL, diclofenazine 1 μg / mL, heparan sulfate glycoprotein 15 μg / mL, difuran ene 1ng / mL, D-menthone 2ng / mL, carbalactone 10μg / mL, IFN-γ5ng / mL, EGF10ng / mL, zinc citrate 12μg / mL.

[0028] The preparation method of the above-mentioned serum-free medium for induced pluripotent stem cells is the same as that in Example 1.

Embodiment 3

[0030] A serum-free medium for induced pluripotent stem cells, consisting of basal medium, adding the following components in the basal medium: insulin, diclofenazine, heparan sulfate glycoprotein, furadienes, D-menthone, Composed of kavalactone, IFN-γ, EGF, and zinc citrate; the dosage of each component in terms of final concentration is as follows: insulin 15 μg / mL, diclofenazine 4 μg / mL, heparan sulfate glycoprotein 20 μg / mL, difuran ene 5ng / mL, D-menthone 7ng / mL, kavalactone 15μg / mL, IFN-γ10ng / mL, EGF15ng / mL, zinc citrate 17μg / mL.

[0031] The preparation method of the above-mentioned serum-free medium for induced pluripotent stem cells is the same as that in Example 1.

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Abstract

The invention discloses a serum-free culture medium for induced pluripotent stem cells. The serum-free culture medium comprises a basic culture medium and the following components added into the basic culture medium: insulin, dichlorophenazine, heparin sulfate glycoprotein, furanodiene, D-menthone, kavalactone, IFN-gamma, EGF and zinc citrate. The dichlorophenazine, the heparin sulfate glycoprotein and the kavalactone which are added into the culture medium have a synergistic effect to regulate and control the mitosis cycle of the induced pluripotent stem cells, accelerate cell metabolism, shorten the cell division cycle and enable the cells to maintain a relatively high proliferation speed. Furandiene and D-menthone are helpful for the induced pluripotent stem cells to maintain proliferation activity and a relatively good growth state, and play a role in ensuring the stability of cell proliferation. IFN-gamma, EGF and zinc citrate provide nutrition supply for cell proliferation, and provide energy for cell proliferation together with the basic culture medium. The invention further provides a preparation method of the serum-free culture medium, the process is simple and easy to operate, and convenience is provided for commercialization of products.

Description

technical field [0001] The invention relates to a serum-free medium, in particular to a serum-free medium for inducing pluripotent stem cells and a preparation method thereof. Background technique [0002] Induced pluripotent stem cells (iPS cells) are transfected with pluripotency genes Sox2, Klf4, OCT4 and c-Myc into somatic cells with retroviruses, and reprogrammed to be able to proliferate and differentiate indefinitely Similar to embryonic stem cells, it can proliferate indefinitely in vitro and in vivo, and can differentiate into almost all types of cells in the human body; at the same time, induced pluripotent stem cells have the advantages of easy extraction, no immune rejection, and no moral and ethical disputes. Therefore, it becomes an ideal seed cell in the research of regenerative medicine treatment. [0003] However, one of the key factors that currently restricts human induced pluripotent stem cells from clinical transformation and drug screening is the lack ...

Claims

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Application Information

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IPC IPC(8): C12N5/10
CPCC12N5/0696C12N2500/30C12N2501/11C12N2501/24C12N2501/33C12N2501/91
Inventor 齐国友
Owner 齐国友
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