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Serum-free medium of stem cell

A serum-free medium and stem cell technology, applied in the field of cell culture medium, can solve the problems of difficult to completely remove serum components, low purity and dryness of stem cells, and affect the quality of final products, so as to inhibit adherent growth and ensure purity and dryness, short cell cycle effect

Active Publication Date: 2015-07-29
钜威细胞(厦门)医学科技有限公司
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0003] 1) The serum is susceptible to contamination by viruses, mycoplasma or other pathogens;
[0004] 2) Quality differences between different batches of serum cause quality differences between final product batches;
[0006] 4) It is difficult to completely remove some serum components, which seriously affects the quality of the final product and poses a safety hazard
However, the above-mentioned serum-free medium is not differentiated for different cells, and there are still some problems in stem cell culture, such as stem cells are prone to differentiation, and the purity and stemness of stem cells are low.

Method used

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  • Serum-free medium of stem cell
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  • Serum-free medium of stem cell

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Embodiment Construction

[0027] The present invention will be further described in detail below using the isolation and culture of adipose-derived mesenchymal stem cells.

[0028] The steps of isolation and culture of adipose-derived mesenchymal stem cells are as follows:

[0029] Obtaining fat tissue: Professional hospitals or beauty institutions obtain fat tissue from the superficial abdominal layer of the patient's abdomen through tumescent liposuction, among which, the patient has passed the screening, that is, HBV antigen, anti-HCV antibody, anti-HIV antibody, and anti-Treponema pallidum antibody , ALT, and mycoplasma detection items were all negative.

[0030] Digestion of adipose tissue: wash the adipose tissue with normal saline, take 10ml of adipose tissue, add an equal volume of 0.4% (v / v) P-type collagenase, mix well, and digest at 37°C and 150rpm for 16min.

[0031] Centrifugal collection: the digested product was filtered through a 150-mesh sieve to collect the initial cell suspension, w...

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Abstract

The invention discloses a serum-free medium of a stem cell, which comprises a basal medium and an additive, wherein the basal medium is DMEM / F12 (Dulbecco Modified Eagle Medium / F12); the additive comprises 2-15%(v / v) of serum replacement, 20-100ug / ml vitamin C, 0.5-10ng / ml stem cell growth factor, 5-20ng / ml human platelet-derived growth factor and 1-5mmol / ml L-glutamine. The serum-free medium of the stem cell for cell culture has the advantages of short cell cycle, strong multiplication capacity, good cell uniformity and high purity, effectively inhibits differentiation of the stem cell and adherence growth of an endothelial cell, ensures purity and dryness of the stem cell and is free from ingredients of animal origin such as fetal calf serum, and the stem cell obtained by the medium is suitable for clinical application.

Description

technical field [0001] The invention relates to a medium for cell culture, in particular to a serum-free medium for stem cells. Background technique [0002] Cell culture is widely used in stem cell cryopreservation. In the traditional culture process, it is usually necessary to add a certain amount of fetal bovine serum, which contains growth factors, hormones, carrier proteins, adhesion factors, trace elements and other nutrients required for cell growth, which can promote cell growth. However, in large-scale production and practical application, the addition of serum has the following disadvantages: [0003] 1) The serum is susceptible to contamination by viruses, mycoplasma or other pathogens; [0004] 2) Quality differences between different batches of serum cause quality differences between final product batches; [0005] 3) The existence of a large amount of serum increases the difficulty of purification in the later stage of production, increases the process flow,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 吴炯
Owner 钜威细胞(厦门)医学科技有限公司
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