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Adipose-derived stem cell serum-free culture medium and preparation method and application thereof

A technology of serum-free medium and adipose stem cells, which is applied in the field of stem cell culture, can solve the problems of enhancing the adhesion performance of adipose stem cells, poor adhesion performance, complex components, etc., so as to facilitate the proliferation, maintain the characteristics of stem cells, and improve the adhesion performance and proliferation rate, the effect of simple preparation method

Active Publication Date: 2017-05-31
广东省科玮智丽生物医药有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Animal serum contains nutrients such as hormones, growth factors, and transfer proteins that can provide adipose-derived stem cells with growth and proliferation, and can promote the attachment of adipose-derived stem cells, but there are many disadvantages: expensive, unstable sources, and large differences between batches , requires a lot of verification work, the composition is not clear, it is not conducive to the separation and purification of vaccines and monoclonal antibodies and other purpose products, and it is easy to be infected by viruses and mycoplasma, etc.
[0005] Chinese patent application CN 103255103 discloses a serum-free fat stem cell culture medium, which adds basic fibroblast growth factor, epidermal growth factor, heparin, L-glutamine, 2-Mercaptoethanol, leukemia inhibitory factor, sodium pyruvate and other components, but no cytokines to promote adhesion, need to use coated culture vessels to improve adhesion performance, and no nutritional ingredients such as albumin, the proliferation is relatively slow slow
[0006] Chinese patent application CN 104877962 discloses a serum-free fat stem cell culture medium, which, on the basis of IMDM medium, also includes recombinant human insulin, recombinant transferrin, vitamin C, human serum albumin, fibroblast Growth factor, platelet-derived growth factor, epidermal growth factor, insulin-like growth factor, transforming growth factor, fibronectin, fetuin, tea polyphenols and stem cell growth factor, through the synergistic effect of fibronectin and fetuin, greatly Enhanced the adhesion performance of adipose-derived stem cells without coating the culture vessel, but the composition is more complex
[0007] At present, there are some commercially available serum-free media for adipose-derived stem cells, such as Gibco, Baitong Biotechnology and other companies' adipose-derived stem cell serum-free media, but there are disadvantages such as high price and unsatisfactory adherence performance. A series of reactions such as the proliferation and differentiation of adipose-derived stem cells affect

Method used

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  • Adipose-derived stem cell serum-free culture medium and preparation method and application thereof
  • Adipose-derived stem cell serum-free culture medium and preparation method and application thereof
  • Adipose-derived stem cell serum-free culture medium and preparation method and application thereof

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Effect test

Embodiment 1

[0027] Embodiment 1 Adipose stem cell serum-free medium

[0028] Adipose stem cell serum-free medium, including DMEM low-glucose medium, also includes transferrin, serum albumin, platelet-derived growth factor, epidermal growth factor, transforming growth factor, β-mercaptoethanol, dihydromyricetin and catechin; The concentration of transferrin was 25 μg / mL, the concentration of serum albumin was 2 mg / mL, the concentration of platelet-derived growth factor was 25 ng / mL, the concentration of epidermal growth factor was 25 ng / mL, and the concentration of transforming growth factor was 25 ng / mL , the concentration of β-mercaptoethanol was 10 μg / mL, the concentration of dihydromyricetin was 25 μg / mL, and the concentration of catechin was 20 μg / mL. In the present invention, the concentration of each component is based on the final volume of the serum-free culture medium of the adipose stem cells.

[0029]Preparation method: take DMEM low-sugar medium, add transferrin, serum albumi...

Embodiment 2

[0030] Example Two Adipose Stem Cell Serum-Free Medium

[0031] Adipose stem cell serum-free medium, including DMEM low-glucose medium, also includes transferrin, serum albumin, platelet-derived growth factor, epidermal growth factor, transforming growth factor, β-mercaptoethanol, dihydromyricetin and catechin; The concentration of transferrin was 40 μg / mL, the concentration of serum albumin was 1 mg / mL, the concentration of platelet-derived growth factor was 30 ng / mL, the concentration of epidermal growth factor was 15 ng / mL, and the concentration of transforming growth factor was 20 ng / mL , the concentration of β-mercaptoethanol was 5 μg / mL, the concentration of dihydromyricetin was 30 μg / mL, and the concentration of catechin was 10 μg / mL.

[0032] Preparation method: take DMEM low-sugar medium, add transferrin, serum albumin, platelet-derived growth factor, epidermal growth factor, transforming growth factor, β-mercaptoethanol, dihydromyricetin and catechin, stir evenly, an...

Embodiment 3

[0033] Example 3 Serum-free Medium for Adipose Stem Cells

[0034] Adipose stem cell serum-free medium, including DMEM low-glucose medium, also includes transferrin, serum albumin, platelet-derived growth factor, epidermal growth factor, transforming growth factor, β-mercaptoethanol, dihydromyricetin and catechin; The concentration of transferrin was 20 μg / mL, the concentration of serum albumin was 3 mg / mL, the concentration of platelet-derived growth factor was 20 ng / mL, the concentration of epidermal growth factor was 20 ng / mL, and the concentration of transforming growth factor was 10 ng / mL , the concentration of β-mercaptoethanol was 20 μg / mL, the concentration of dihydromyricetin was 10 μg / mL, and the concentration of catechin was 25 μg / mL.

[0035] Preparation method: take DMEM low-sugar medium, add transferrin, serum albumin, platelet-derived growth factor, epidermal growth factor, transforming growth factor, β-mercaptoethanol, dihydromyricetin and catechin, stir evenly...

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Abstract

The invention provides an adipose-derived stem cell serum-free culture medium which comprises a DMEM low-sugar culture medium and further comprises transferrin, serum albumin, a platelet-derived growth factor, an epidermal growth factor, a transforming growth factor, beta-mercaptoethanol, dihydroarbutin, and catechin. The adipose-derived stem cell serum-free culture medium belongs to the technical field of cell culture. The adipose-derived stem cell serum-free culture medium can remarkably improve the adherence performance and multiplication rate of adipose-derived stem cells and facilitates multiplication and property keeping of the adipose-derived stem cells, the ingredients of the culture medium are simpler, and the cost is lower.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, and in particular relates to a serum-free culture solution of adipose stem cells and its preparation method and application. Background technique [0002] Adipose-derived stem cells (ADSCs) are a kind of mesenchymal stem cells with multidirectional differentiation potential isolated from adipose tissue by Zuk et al. in 2001. Adipose stem cells can proliferate stably in vitro, and are rich in sources, convenient to obtain materials, and have a large amount of cells to obtain. They are suitable for large-scale culture and have high clinical application value. They have been widely used in the fields of cell engineering, tissue engineering, and regenerative medicine. [0003] Adipogenic stem cells adhere slowly, and it is usually necessary to add animal serum or other growth factors that promote adherence to the culture medium to promote the adherence and proliferation of adipose stem cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/24C12N2500/30C12N2500/34C12N2500/44C12N2500/90C12N2501/11C12N2501/135C12N2501/148C12N2501/15C12N2501/998
Inventor 陈松彬倪彦艳廖丽影
Owner 广东省科玮智丽生物医药有限公司
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