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Culture medium and culture method for culturing mesenchymal stem cells

A mesenchymal stem cell and culture medium technology, which is applied in the field of culture medium for mesenchymal stem cells, can solve the problems of inconvenient activation of mesenchymal stem cells, low cell viability and secretion ability, and affect the speed of cell proliferation, etc., to delay mesenchymal stem cells. The effects of aging and functional decline of stem cells, improving activity and secretion capacity, and increasing cell proliferation speed

Pending Publication Date: 2021-12-28
上海泽充生物技术有限公司
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Problems solved by technology

[0003] At present, there are still some problems in the existing medium and culture methods for culturing mesenchymal stem cells: 1. The abnormal expression of microRNA in the aging process of mesenchymal stem cells, resulting in the decline of BMSC function; 2. It is inconvenient to activate mesenchymal stem cells The vitality potential of the cells, the activity and secretion capacity of the cells are low, which affects the cell proliferation speed, so we propose a medium and a culture method for culturing mesenchymal stem cells

Method used

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  • Culture medium and culture method for culturing mesenchymal stem cells
  • Culture medium and culture method for culturing mesenchymal stem cells
  • Culture medium and culture method for culturing mesenchymal stem cells

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Embodiment 1

[0036] see Figure 1-Figure 2 , the present invention provides a technical solution: a medium for culturing mesenchymal stem cells, including the following component raw materials: serum 5 μl / ml, miR-705antagomir 8ng / mL, ascorbic acid 30mg / L, honeysuckle powder 2ng / mL , luteolin 15μg / mL, epidermal growth factor 8ng / mL, penicillin 40U / mL, streptomycin 70μg / mL, vitamin C25μg / mL and cell basal medium.

[0037] In this embodiment, preferably, the cell basal medium is DMEM, and the serum is fetal bovine serum.

[0038] In this embodiment, preferably, the preparation method of the medium for culturing mesenchymal stem cells comprises the following steps:

[0039] S1. Preparation of raw materials: Prepare raw materials according to the original composition, serum 5 μl / ml, miR-705antagomir 8ng / mL, ascorbic acid 30mg / L, honeysuckle powder 2ng / mL, luteolin 15μg / mL, epidermal growth factor 8ng / mL, penicillin 40U / mL, streptomycin 70μg / mL, vitamin C25μg / mL and cell basal medium;

[004...

Embodiment 2

[0059] see Figure 1-Figure 2 , the present invention provides a technical solution: a culture medium for culturing mesenchymal stem cells, including the following component raw materials: serum 50 μl / ml, miR-705antagomir 16ng / mL, ascorbic acid 60mg / L, honeysuckle powder 5ng / mL , luteolin 30μg / mL, epidermal growth factor 20ng / mL, penicillin 60U / mL, streptomycin 80μg / mL, vitamin C 100μg / mL and cell basal medium.

[0060] In this embodiment, preferably, the cell basal medium is α-MEM, and the serum is human serum.

[0061] In this embodiment, preferably, the preparation method of the medium for culturing mesenchymal stem cells comprises the following steps:

[0062] S1. Preparation of raw materials: prepare raw materials according to the original composition, serum 50μl / ml, miR-705antagomir8-16ng / mL, ascorbic acid 60mg / L, honeysuckle powder 5ng / mL, luteolin 30μg / mL, epidermal growth factor 20ng / mL, Penicillin 60U / mL, streptomycin 80μg / mL, vitamin C100μg / mL and cell basal mediu...

Embodiment 3

[0082] see Figure 1-Figure 2 , the present invention provides a technical solution: a medium for culturing mesenchymal stem cells, including the following component raw materials: serum 20 μl / ml, miR-705antagomir 12ng / mL, ascorbic acid 40mg / L, honeysuckle powder 3ng / mL , luteolin 20μg / mL, epidermal growth factor 12ng / mL, penicillin 50U / mL, streptomycin 75μg / mL, vitamin C 50μg / mL and cell basal medium.

[0083] In this embodiment, preferably, the cell basal medium is DMEM / F12, and the serum is fetal bovine serum.

[0084] In this embodiment, preferably, the preparation method of the medium for culturing mesenchymal stem cells comprises the following steps:

[0085] S1. Preparation of raw materials: prepare raw materials according to the original composition, serum 20μl / ml, miR-705antagomir12ng / mL, ascorbic acid 40mg / L, honeysuckle powder 3ng / mL, luteolin 20μg / mL, epidermal growth factor 12ng / mL, penicillin 50U / mL, streptomycin 75μg / mL, vitamin C 50μg / mL and cell basal mediu...

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Abstract

The invention discloses a culture medium for culturing mesenchymal stem cells. The culture medium is prepared from the following component raw materials: 5 to 50 [mu] l / ml of serum, 8 to 16 ng / mL of miR-705antagomir, 30 to 60 mg / L of ascorbic acid, 2 to 5 ng / mL of honeysuckle flower powder, 15 to 30 [mu] g / mL of luteolin, 8 to 20 ng / mL of epidermal growth factors, 40 to 60 U / mL of penicillin, 70 to 80 [mu] g / mL of streptomycin, 25 to 100 [mu] g / mL of vitamin C and a cell basal culture medium. The invention further discloses a culture method for culturing the mesenchymal stem cells. The culture method comprises the following steps: step 1, preparing an incubator; step 2, obtaining bone marrow mesenchymal stem cells; and step 3, culturing the bone marrow mesenchymal stem cells. Aiming at microRNA abnormal expression in the aging process of the mesenchymal stem cells, a specific microRNA inhibitor antagomir is adopted, a transfection reagent and a carrier are not needed, and the microRNA inhibitor antagomir directly enters the cells, so that cell expression is safely and efficiently regulated, miR-705 is reduced, the aging and function reduction of the mesenchymal stem cells caused by long-term in-vitro passage amplification are delayed, and a foundation is established for clinical application of the BMSC.

Description

technical field [0001] The invention belongs to the technical field of culture medium, and in particular relates to a culture medium and a culture method for culturing mesenchymal stem cells. Background technique [0002] Culture medium refers to a nutrient matrix prepared by combining different nutrients for the growth and reproduction of microorganisms, plants or animals (or tissues). Generally contain carbohydrates, nitrogenous substances, inorganic salts (including trace elements), vitamins and water and other major categories of substances. The medium is not only the basic material for providing nutrition and promoting cell proliferation, but also the living environment for cell growth and reproduction. There are many types of medium, which can be divided into natural medium, synthetic medium, and semi-synthetic medium according to the source of the prepared raw materials; solid medium, liquid medium, and semi-solid medium according to the physical state; It can be di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2500/76C12N2501/11C12N2500/38C12N2500/05C12N2500/30C12N2501/599C12N2501/00
Inventor 李一佳
Owner 上海泽充生物技术有限公司
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