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Serum-free medium for adipose-derived mesenchymal stem cells and its preparation method and application
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A technology of serum-free medium and mesenchymal stem cells, applied in the direction of cell culture active agents, biochemical equipment and methods, culture process, etc., to achieve stable performance, increase the expansion rate, and improve the survival rate
Active Publication Date: 2022-02-18
BAOXIN ASIA PACIFIC BIOTECH SHENZHEN CO LTD
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[0005] The purpose of this application is to provide a serum-free medium for adipose-derived mesenchymal stem cells and its preparation method, as well as an application of a serum-free medium for adipose-derived mesenchymal stem cells, aiming to solve the problem of existing adipose-derived mesenchymal stem cells to a certain extent. The problem of using serum-containing medium as a medium for stem cell culture
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[0067] The second aspect of the embodiment of the present application provides a method for preparing a serum-free medium of adipose-derived mesenchymal stem cells, comprising the following steps:
[0068] S10. Obtain transferrin, recombinant human insulin, fibroblast growth factor, fibronectin, astragaloside IV, Codonopsis pilosula extract and aloe extract respectively;
[0069] S20. Mix transferrin, recombinant human insulin, fibroblast growth factor, fibronectin, astragaloside IV, Codonopsis pilosula extract and aloe extract with DMEM / F12 medium to obtain adipose-derived mesenchymal stem cells without serum medium.
[0070] The preparation method of the serum-free medium of adipose-derived mesenchymal stem cells provided in the second aspect of the embodiment of the present application is to obtain transferrin, recombinant human insulin, fibroblast growth factor, fibronectin, astragaloside IV, and Codonopsis pilosula extract respectively After being mixed with aloe vera ex...
Embodiment 1
[0125] A kind of adipose-derived mesenchymal stem cell has / serum-free medium, comprises DMEM / F12 medium, the transferrin of final concentration is 8 μ g / mL, the recombinant human insulin of 10 μ g / mL, the fibroblast growth factor of 10 ng / mL, 50μg / mL of fibronectin, the concentrations of astragaloside IV were 6mg / l, 12mg / l, 24mg / l, 48mg / l and 96mg / l.
Embodiment 2
[0127] A kind of adipose-derived mesenchymal stem cell has / serum-free culture medium, comprises DMEM / F12 medium, the transferrin of final concentration is 15 μ g / mL, the recombinant human insulin of 20 μ g / mL, the fibroblast growth factor of 20 ng / mL, 70μg / mL fibronectin, the concentration of Codonopsis pilosula extract is 5mg / l, 10mg / l, 20mg / l, 40mg / l and 60mg / l respectively.
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Abstract
The application belongs to the technical field of cell culture medium, and in particular relates to a serum-free medium for adipose-derived mesenchymal stem cells and a preparation method thereof, as well as an application of a serum-free medium for adipose-derived mesenchymal stem cells. Wherein, the serum-free medium of adipose-derived mesenchymal stem cells includes basal medium and supplements; wherein, the basal medium comprises DMEM / F12 medium; the supplements include transferrin, recombinant human insulin, fibroblast Cell Growth Factor, Fibronectin, Astragaloside IV, Codonopsis Extract and Aloe Vera Extract. The serum-free medium of this application is used to cultivate adipose-derived mesenchymal stem cells. Through the synergistic effect of various components, it can not only promote the growth and proliferation of adipose-derived stem cells, but also reduce cell damage and improve the survival rate of cells, thereby improving the adipose-derived stem cells. In vitro expansion capability. Moreover, there is no heterologous pollution, no potential safety hazards causing rejection, and the performance of the culture medium is stable and the quality is reliable.
Description
technical field [0001] The application belongs to the technical field of cell culture medium, and in particular relates to a serum-free medium for adipose-derived mesenchymal stem cells and a preparation method thereof, as well as an application of a serum-free medium for adipose-derived mesenchymal stem cells. Background technique [0002] Stem cells are a type of pluripotent cells with self-renewal and multi-directional differentiation capabilities. Mesenchymal stem cells are an important member of the stem cell family, derived from the mesoderm in the early stage of development, and can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, Various tissue cells such as endothelium are ideal seed cells for regenerative medicine. Mesenchymal stem cells exist in bone marrow, umbilical cord blood and umbilical cord tissue, placental tissue, adipose tissue and other tissues. When cultured in vitro, it is easy to adhere to the wall an...
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