Serum-free medium for placenta-derived mesenchymal stem cells and preparation method thereof

A serum-free medium and stem cell technology, applied in the field of stem cells, can solve problems such as poor safety and unsatisfactory cell proliferation rate, and achieve the effects of high safety, excellent cell performance, and good cell morphology

Active Publication Date: 2015-12-02
广东美赛尔细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the defects of poor safety and unsatisfactory cell proliferation rate in the serum-free medium of placental mesenchymal stem cells in the prior art, the purpose of the present invention is t

Method used

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  • Serum-free medium for placenta-derived mesenchymal stem cells and preparation method thereof
  • Serum-free medium for placenta-derived mesenchymal stem cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, the influence of the culture medium containing different concentrations of magnolol on cell proliferation

[0023] The inventor conducted a large number of tests to screen and obtain the serum-free medium formula of placental mesenchymal stem cells provided by the present invention, and further investigated the influence of the concentration of magnolol.

[0024] Dissolve magnolol powder in dimethyl sulfoxide into 100 mM magnolol stock solution, and dilute the magnolol stock solution into 35 μM, 30 μM, 25 μM, 12.5 μM, 6.25 μM, 3.13 μM with high-glucose DMEM basal medium respectively , 1.56μM, 0.78μM, 0.39μM drug solutions.

[0025] Placental mesenchymal stem cells of P1 generation were cultured at 10 4 Cells / mL were inoculated in 96-well plates, cultured in serum-free medium containing different concentrations of magnolol-containing placental mesenchymal stem cells, 100 μL per well, and the density of placental mesenchymal stem cells without magnolol S...

Embodiment 2

[0026] Embodiment two, Serum-free medium for placental mesenchymal stem cells

[0027] Serum-free medium for placental mesenchymal stem cells, including high-glucose DMEM basal medium, also includes the following components and their concentrations: vitamin H1ug / ml, glutathione 5ug / ml, recombinant human insulin 3mg / ml, human Serum albumin 0.5mg / ml, transferrin 2ug / ml, fibroblast growth factor 0.5ng / ml, epidermal growth factor 0.5ng / ml, stem cell growth factor 0.5ng / ml, human stem cell factor 0.5ng / ml and thick Parkol 6uM.

[0028] Preparation method: Add vitamin H, glutathione, recombinant human insulin, human serum albumin, transferrin, fibroblast growth factor, epidermal growth factor, stem cell growth factor, Human stem cell factor and magnolol, mixed evenly, passed through a membrane to remove bacteria.

Embodiment 3

[0029] Embodiment three, Serum-free medium for placental mesenchymal stem cells

[0030] Serum-free medium for placental mesenchymal stem cells, including high-glucose DMEM basal medium, also includes the following components and their concentrations: vitamin H5ug / ml, glutathione 10ug / ml, recombinant human insulin 6.5mg / ml, Human serum albumin 1mg / ml, transferrin 6ug / ml, fibroblast growth factor 10ng / ml, epidermal growth factor 5ng / ml, stem cell growth factor 5ng / ml, human stem cell factor 5ng / ml and magnolol 25uM.

[0031] The preparation method is similar to Example 2.

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Abstract

The invention belongs to the technical field of stem cells and relates to a serum-free medium for placenta-derived mesenchymal stem cells and a preparation method thereof. The serum-free medium comprises a DMEM basic medium and further comprises vitamin H, glutathione, recombinant human insulin, human serum albumin, transferrin, fibroblast growth factors, epidermal growth factors, stem cell growth factors, Human stem cell factors and magnolol. The serum-free medium provided by the invention has the advantages of high safety and capability for significant improvement of proliferative capability of placenta-derived mesenchymal stem cells, the serum-free medium is capable of well keeping cell forms and stem cell characteristics of the placenta-derived mesenchymal stem cells and is convenient to popularize and apply.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a serum-free culture medium of placental mesenchymal stem cells and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) is another adult stem cell in bone marrow besides hematopoietic stem cells, widely distributed in various tissues such as bone marrow, liver and fat in animals. MSCs are a kind of cells with strong self-renewal ability and multi-directional differentiation potential, and MSCs are also an important immune regulatory cell. MSCs exhibit a strong inhibitory effect on the immune system after stimulation by inflammatory cytokines, which can reduce immune rejection in vivo and prolong the survival time of grafts. Therefore, mesenchymal stem cells have broad application prospects in bone and cartilage in tissue engineering, seed cells for myocardial construction, hematopoietic stem cells, and organ transpla...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 王一飞廖晓凤王巧利
Owner 广东美赛尔细胞生物科技有限公司
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