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Stem cell serum-free culture medium as well as preparation method and application thereof

A serum-free medium and stem cell culture technology, applied in the direction of cell culture active agents, biochemical equipment and methods, culture process, etc., can solve the problems of protein molecular cell internalization, contamination by pathogens, interference with therapeutic effect, etc., to achieve composition Simple, high safety, low cost effect

Pending Publication Date: 2022-03-22
KYBIOSTEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the limitations of fetal bovine serum gradually emerged in the process of use: fetal bovine serum is obtained from the blood of fetal bovines, and there is a high possibility of being contaminated by pathogens during the extraction process. During the culture process of stromal stem cells, these infectious agents may be transmitted to the cells; the phagocytosis of the cells will also internalize the protein molecules in the fetal bovine serum, which cannot be removed by washing, and clinical treatment usually requires multiple infusions in small doses. Injecting cells, the residual foreign protein directly entering the human body will cause the body's immune rejection, which will interfere with the therapeutic effect to a certain extent; at the same time, there are risks to product technicians and medical personnel (when handling products); fetal bovine serum needs to be in- Refrigerated storage at 20°C, and there are problems such as large differences between batches

Method used

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  • Stem cell serum-free culture medium as well as preparation method and application thereof
  • Stem cell serum-free culture medium as well as preparation method and application thereof
  • Stem cell serum-free culture medium as well as preparation method and application thereof

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preparation example Construction

[0069] In a second aspect, the present invention provides a method for preparing a stem cell serum-free medium, comprising the following steps:

[0070] Glucose, mannitol, glycine and polyethylene glycol are dissolved in basal medium, and then mixed with insulin solution and metformin solution to prepare stem cell serum-free medium;

[0071] Preferably, the basal medium contains antibiotics.

[0072] It should be noted that the order of mixing the basal medium with the insulin solution and the metformin solution is not specifically limited in the present invention, for example, the basal medium can be mixed with the insulin solution and then mixed with the metformin solution, or the basal medium can be mixed now It is mixed with metformin solution and then mixed with insulin solution, and insulin solution and metformin solution can also be mixed and then mixed with basal medium.

[0073] The preparation method of the stem cell serum-free medium provided by the invention is si...

Embodiment 1

[0085] A serum-free medium for culturing stem cells, including: DMEM containing 1w / v% penicillin-streptomycin double antibody and 100μM metformin, 10μg / mL insulin, 1w / v% mannitol, 1w / v% poly Ethylene glycol, 1w / v% glycine, 5w / v% glucose.

[0086] The preparation method is as follows:

[0087] Dissolve mannitol, glycine, and polyethylene glycol in DMEM medium containing 1% penicillin-streptomycin double antibody, filter through a 0.22 μm sterile filter; dissolve insulin with 10 mM hydrochloric acid to a concentration of 1 mg / mL concentration, use a 0.22 sterile filter to filter; use PBS to dissolve metformin to a concentration of 10mM, use a 0.22 sterile filter to filter; then add insulin solution and metformin solution to the culture medium according to the required final concentration middle.

Embodiment 2

[0089] A serum-free medium for culturing stem cells, including: DMEM containing 1w / v% penicillin-streptomycin double antibody and 100μM metformin, 100μg / mL insulin, 1w / v% mannitol, 1w / v% poly Ethylene glycol, 1w / v% glycine, 5w / v% glucose.

[0090] The preparation method is the same as in Example 1.

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Abstract

The invention provides a stem cell serum-free culture medium as well as a preparation method and application thereof, and relates to the technical field of biology. The stem cell serum-free culture medium provided by the invention comprises a basic culture medium, and metformin, insulin, mannitol, glycine, polyethylene glycol and glucose with specific concentrations, and the research of the inventor finds that the metformin has the capability of improving stem cell proliferation; metformin is used in cooperation with insulin, mannitol, glycine, polyethylene glycol and glucose, serum can be replaced to achieve stem cell culture, the stem cell serum-free culture medium is simple in composition and high in safety, good cell viability and proliferation rate can be kept, the osteogenesis and adipogenesis differentiation inducing capacity of stem cells is improved, and the stem cell serum-free culture medium is suitable for being used in the field of stem cell culture. The problems caused by serum use are effectively avoided, and the method can be applied to stem cell culture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a serum-free culture medium for stem cells and its preparation method and application. Background technique [0002] Mesenchymal Stem Cells (MSCs) are a kind of pluripotent stem cells, which have all the common characteristics of stem cells, that is, self-replication ability and multi-directional differentiation ability, and can be obtained from a variety of tissues, including umbilical cord blood, bone marrow, adipose tissue , periodontal ligament, pulp and gingiva. Stem cells from human exfoliated deciduous teeth (SHED) belong to ectodermal mesenchymal stem cells and are an ideal source of stem cells. Through directional induction, dental pulp stem cells of deciduous teeth can differentiate into bone cells, liver cells in vitro and in vivo. The ability of stem cells, muscle cells, nerve cells and skin cells has significant advantages and good application prospects in the treatment...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/90C12N2500/35C12N2500/34C12N2500/30C12N2500/32C12N2501/33
Inventor 邓世文雷童郑慧婷陈虹羽杜宏武
Owner KYBIOSTEM CO LTD
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