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Serum-free culture medium and stem cell culture method

A serum-free medium, basal medium technology, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve problems such as pollution, and achieve the effect of improving repeatability, consistent quality and low protein content

Inactive Publication Date: 2015-11-25
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a non-toxic and non-polluting serum-free medium in view of the defects of certain toxicity and pathogen contamination in the prior art of using animal serum to cultivate cells

Method used

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  • Serum-free culture medium and stem cell culture method

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Embodiment 1

[0038] In this embodiment, the components of the serum-free medium and the concentration of each component in the serum-free medium are respectively:

[0039] DMEM / F12 medium: 500mL;

[0040] Insulin: 10μg / ml;

[0041] Transferrin: 10μg / ml;

[0042] Ferric citrate: 10μg / ml

[0043] Sodium selenite: 20μg / ml;

[0044] Fibronectin: 2 μg / ml;

[0045] Glucagon: 3 μg / ml;

[0046] Hepatocyte growth factor: 10ng / ml;

[0047] Penicillin streptomycin: 50U / ml.

Embodiment 2

[0049] In this embodiment, the components of the serum-free medium and the concentration of each component in the serum-free medium are respectively:

[0050] DMEM / F12 medium: 500mL;

[0051] Insulin: 6 μg / ml;

[0052] Transferrin: 7μg / ml;

[0053] Sodium selenite: 15μg / ml;

[0054] Epidermal growth factor: 15ng / ml;

[0055] Hepatocyte growth factor: 15ng / ml;

[0056] Fibronectin: 1.5 μg / ml;

[0057] Glucagon: 5 μg / ml;

[0058] HEPES: 15mmol / L;

[0059] Penicillin streptomycin: l20U / ml.

Embodiment 3

[0061] In this embodiment, the components of the serum-free medium and the concentration of each component in the serum-free medium are respectively:

[0062] DMEM / F12 medium: 500mL;

[0063] Insulin: 5 μg / ml;

[0064] Transferrin: 5μg / ml;

[0065] Sodium selenite: 10μg / ml;

[0066] Epidermal growth factor: 20ng / ml;

[0067] Hepatocyte growth factor: 20ng / ml;

[0068] Fibronectin: 1 μg / ml;

[0069] Glucagon: 4 μg / ml;

[0070] HEPES: l0mmol / l;

[0071] Penicillin streptomycin: l00U / ml;

[0072] Dexamethasone: 1 nmol / ml.

[0073] In order to further verify that the serum-free medium provided by the present invention has an outstanding and significant effect, the following specific experiments will be further described in detail.

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Abstract

The invention relates to a serum-free culture medium which comprises a basal culture medium, insulin, iron-containing food additive, sodium arsenite, fibronectin, glucagon, hepatocyte growth factor and myllicin. In addition, the invention further discloses a stem cell culture method. The stem cell culture method includes the following steps: acquiring stem cells; adopting the serum-free culture medium for in vitro culture. By the serum-free culture medium, the problem of difficulty in cell purifying due to pollution of pathogens carried by a serum culture medium can be avoided, stability of cell production can be improved, cell proliferation amount can be increased, and cell activity can be enhanced.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a serum-free medium and a method for culturing stem cells. Background technique [0002] At present, a certain proportion of animal serum is added to the conventional culture system of stem cells, and fetal bovine serum or newborn calf serum is more commonly used. Serum is an extremely complex mixture composed of many biomolecules of different sizes. Its main role in culturing cells in vitro is to provide growth factors, hormones, binding proteins, and provide protection. But it also contains some inhibitory factors or toxic substances that are not conducive to cell growth, and have potential cytotoxic effects. A large number of complex proteins in serum bring difficulties to the standardization of cell culture, and also to the separation and purification of cell culture expression products. Moreover, animal serum may contain known or unknown pathogens carried by ani...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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