Serum-free culture medium of placental mesenchymal stem cells

A serum-free medium and quality stem cell technology, applied in the field of stem cells, can solve the problems of unclear composition, content and its mechanism of action, adverse effects on cell growth, and large differences in serum quality, so as to eliminate pathogen contamination and shorten culture time. , the effect of clear components

Active Publication Date: 2020-08-28
广州同康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional culture method of mesenchymal stem cells is to use culture medium containing animal serum, mainly to provide nutrients such as hormones, growth factors, transport proteins, etc. for the growth and reproduction of mesenchymal stem cells. However, serum is a very complex There may be as many as several hundred ingredients, and the exact ingredients, content and mechanism of action are still unclear. The quality of serum varies greatly between batches, which is greatly affected by the source of raw materials of the manufacturer. It is very difficult to ensure the consistency of each batch, which is not conducive to the standardization of laboratory research or product production. The serum may contain some unknown or undetected harmful microorganisms such as m

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the influence that the culture medium that contains different concentrations of Pluronic F68 has on cell proliferation

[0026] Take Pluronic F68 and add it to normal saline to obtain 100mM Pluronic F68 mother solution, and use DMEM low-sugar medium to dilute the Pluronic F68 mother solution to 5 μg / mL, 10 μg / mL, 15 μg / mL, 18 μg / mL, respectively. mL, 20 μg / mL drug solution, and then add an equal amount of lignans at the same concentration (6 μg / mL) to the drug solution.

[0027] Placental mesenchymal stem cells of P1 generation were cultured at 10 4 Cells / mL were inoculated in 96-well plates, and culture media containing different concentrations of Pluronic F68 were used, 100 μL per well, for the placental mesenchymal stem cells without Pluronic F68 and lignans. Serum culture medium was the blank group, placed at 37°C, 5% CO 2 After 6h, 8h and 12h respectively, the absorbance value of each well was measured by the MTT detection method, and the absorbance...

Embodiment 2

[0034] Embodiment 2, placental mesenchymal stem cell serum-free medium of the present invention

[0035]Placental mesenchymal stem cell serum-free medium of the present invention comprises DMEM low-sugar medium and supplement, and supplement comprises the component of following concentration: human serum albumin 1mg / mL, transferrin 5μg / mL, epidermal growth factor 5ng / mL mL, transforming growth factor-β 10 ng / mL, platelet growth factor 10 ng / mL, reduced glutathione 1 μg / mL, pluronic F68 10 μg / mL and lignans 5 μg / mL.

[0036] The preparation method of placental mesenchymal stem cell serum-free medium of the present invention comprises the following steps: adding human serum albumin, transferrin, epidermal growth factor, transforming growth factor-beta, platelet growth factor, Reduced glutathione, Pluronic F68 and lignans were stirred evenly at a stirring speed of 800rpm / min for 20min, and filtered through a 0.1μm filter membrane to obtain placental mesenchymal stem cells Serum-...

Embodiment 3

[0038] Embodiment 3, placental mesenchymal stem cell serum-free medium of the present invention

[0039] The serum-free medium for placental mesenchymal stem cells of the present invention includes DMEM low-sugar medium and supplements, and the supplements include components at the following concentrations: human serum albumin 3 mg / mL, transferrin 6 μg / mL, epidermal growth factor 7 ng / mL mL, transforming growth factor-β 20ng / mL, platelet growth factor 15ng / mL, reduced glutathione 2μg / mL, pluronic F68 15μg / mL and lignans 6μg / mL.

[0040] The preparation method of placental mesenchymal stem cell serum-free medium of the present invention comprises the following steps: adding human serum albumin, transferrin, epidermal growth factor, transforming growth factor-beta, platelet growth factor, Reduced glutathione, Pluronic F68 and lignans were stirred evenly at a stirring speed of 900rpm / min for 25min, and filtered through a 0.15μm filter membrane to obtain placental mesenchymal stem...

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Abstract

The invention relates to the technical field of stem cells, in particular to a serum-free culture medium of placental mesenchymal stem cells. The serum-free culture medium of the placental mesenchymalstem cells comprises a DMEM low-sugar culture medium and a supplement, wherein the supplement comprises the following components: human serum albumin with a concentration of 1-5 mg/mL, transferrin with a concentration of 5-10 [mu]g/mL, an epidermal growth factor with a concentration of 5-10 ng/mL, a transforming growth factor with a concentration of 10-30 ng/mL, a platelet-derived growth factor with a concentration of 10-25 ng/mL, glutathione with a concentration of 1-5 [mu]g/mL, Pluronic F68 with a concentration of 10-20 [mu]g/mL and lignans with a concentration of 5-10 [mu]g/mL. The produced serum-free culture medium can significantly improve the proliferation capability of the placental mesenchymal stem cells, a survival rate of the placental mesenchymal stem cells is high, the culturecost is low, the culture time of the cells is shortened, the application range is wide, and the serum-free culture medium of the placental mesenchymal stem cells can be used for industrial production.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a serum-free culture medium for placental mesenchymal stem cells. Background technique [0002] Placental mesenchymal stem cells (MSCs) are a kind of pluripotent stem cells, pluripotent cells with strong proliferation ability and multilineage differentiation potential. MSC has the ability to differentiate into various cells such as epithelial stem cells, neural stem cells, liver stem cells, muscle cells, osteoblasts, chondrocytes, and stromal cells under suitable in vivo or in vitro environments. MSC has an immunoregulatory effect. Through the negative immune regulatory function, it can inhibit the body's hyperactive immune response and restore the balance of the body's immune function, so it can be used to treat immune rejection after hematopoietic stem cell transplantation and Crohn's disease, lupus erythematosus, and scleroderma. diseases and other autoimmune diseases. MS...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N5/0775
CPCC12N5/0605C12N5/0668C12N2500/24C12N2500/90C12N2501/11C12N2501/135C12N2501/15C12N2501/30C12N2501/998C12N2501/999
Inventor 张印
Owner 广州同康生物科技有限公司
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