Premixed liquid culture medium capable of being stored at 4 DEG C for long time
By adding glycerol and antioxidant vitamin C to the culture medium, a protective film is formed and the pH value is stabilized, which solves the problem of nutrient loss in the culture medium at low temperatures and enables long-term preservation and maintenance of cell activity at 4°C.
Patent Information
- Application Number
- CN202511198346.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-26
- Publication Date
- 2025-11-18
AI Technical Summary
Existing culture media are prone to nutrient loss at low temperatures, which affects the growth and reproduction of plant and animal cells and makes them difficult to preserve for long periods.
By adding glycerol to the culture medium to form a protective film and using antioxidants such as vitamin C to prevent the membrane structure from being oxidized and damaged, while adjusting the ratio of growth factors and slow-release agents, the culture medium can be stored for a long time at 4°C without affecting cell activity.
This method enables long-term preservation of the culture medium at 4°C while maintaining the normal function and activity of the cells, thus ensuring the growth and reproduction of plant and animal cells.
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of culture medium, in particular to a premixed liquid culture medium capable of long-term storage at 4 DEG C. BACKGROUND
[0002] Culture medium is a nutrient medium artificially prepared for the growth and reproduction of microorganisms, plants or animal cells At present, when culturing animal and plant cells, culture medium is generally used to provide nutrients for animal and plant cells, but the culture medium needs to be stored at room temperature during use, and the nutrients in the culture medium are prone to loss at low temperature, which is not convenient for long-term storage, and thus affects the growth and reproduction of plant or animal cells during the culturing process, and thus needs to be improved. SUMMARY
[0003] The present application aims to provide a premixed liquid culture medium capable of long-term storage at 4 DEG C, so as to solve the problems in the background art.
[0004] To achieve the above-mentioned purpose, the present application provides the following technical scheme: A premixed liquid culture medium capable of long-term storage at 4 DEG C, comprising a basic culture medium and special additives, the basic culture medium comprising a carbon source, a nitrogen source, inorganic salts and growth factors; the special additives comprising growth regulators and slow-release agents, and further comprising antioxidants and directional differentiation inducers.
[0005] Further, the basic culture medium and the special additives comprise the following mass fractions of raw materials: the carbon source comprises 1.5-2.5% of glucose, the nitrogen source comprises 0.8-1.2% of proteose peptone, the inorganic salts comprise 0.05-0.15% of potassium dihydrogen phosphate and 0.02-0.08% of magnesium sulfate; the growth factors comprise 3-4% of serum, 2-6% of growth factor supplements, 4-7% of basic fibroblast growth factor and 3-6% of epidermal growth factor; the growth regulator comprises 1-3% of antibiotics; the slow-release agent comprises 2-8% of glycerol; and the special additives further comprise 5-15% of antioxidants and 0.3-0.8% of directional differentiation inducers.
[0006] Further, the basic medium and special additives include the following mass fractions of raw materials: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salt including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factor including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, epidermal growth factor 3%-6%; growth regulator including antibiotic 1%-3%; slow-release agent including glycerol 6%-10%; special additives also including antioxidant 5%-15%, directional differentiation inducer 0.3%-0.8%.
[0007] Further, the basic medium and special additives include the following mass fractions of raw materials: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salt including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factor including serum 3%-4%, growth factor supplement 5%-8%, basic fibroblast growth factor 6%-10%, epidermal growth factor 4%-8%; growth regulator including antibiotic 1%-3%; slow-release agent including glycerol 6%-10%; special additives also including antioxidant 5%-15%, directional differentiation inducer 0.3%-0.8%.
[0008] Further, the basic medium and special additives include the following mass fractions of raw materials: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salt including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factor including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, epidermal growth factor 3%-6%; growth regulator including antibiotic 3%-6%; slow-release agent including glycerol 6%-10%; special additives also including antioxidant 5%-15%, directional differentiation inducer 0.3%-0.8%.
[0009] Further, the basic medium and special additives include the following mass fractions of raw materials: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salt including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factor including serum 3%-4%, growth factor supplement 5%-8%, basic fibroblast growth factor 6%-10%, epidermal growth factor 4%-8%; growth regulator including antibiotic 3%-6%; slow-release agent including glycerol 6%-10%; special additives also including antioxidant 5%-15%, directional differentiation inducer 0.3%-0.8%.
[0010] Furthermore, antioxidants include vitamin C.
[0011] Furthermore, the directional differentiation inducer includes dexamethasone.
[0012] Furthermore, serum includes fetal bovine serum.
[0013] Furthermore, antioxidants also include sodium pyruvate.
[0014] Compared with the prior art, the beneficial effects of the present invention are: 1. In this invention, by adding glycerol, the glycerol can form a protective film on the culture medium, and vitamin C can prevent the membrane structure from being oxidized and damaged, thus ensuring normal cell function. With other components unchanged, the slow-release agent is removed. Experiments show that the culture medium containing the slow-release agent can be stored for a long time at 4°C without affecting the activity of plant and animal cells. Detailed Implementation
[0015] To further understand the content of this invention, the invention will be described in detail with reference to the embodiments. It should be understood that the embodiments are merely illustrative and not limiting of the invention. Example 1
[0016] This embodiment provides a premixed liquid culture medium that can be stored at 4°C for an extended period. The medium includes a basal culture medium and special additives. The basal culture medium includes a carbon source, a nitrogen source, inorganic salts, and growth factors. The special additives include growth regulators and slow-release agents, as well as antioxidants and directional differentiation inducers. Example 2
[0017] In this embodiment, the basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth regulators including antibiotics 1%-3%; sustained-release agents including glycerol 2%-8%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%, with the antioxidant including vitamin C and the main directional differentiation inducer including dexamethasone.
[0018] In actual use, the basic culture medium and special additives are added to 150ml of water in sequence to form the culture medium. By adding glycerol, the glycerol can form a protective film on the culture medium, and vitamin C can prevent the membrane structure from being oxidized and damaged, thus ensuring normal cell function. With other components unchanged, the slow-release agent is removed. Experiments show that the culture medium containing the slow-release agent can be stored for a long time at 4°C without affecting the activity of plant and animal cells. Replacing vitamin C with sodium pyruvate allows sodium pyruvate to metabolize into bicarbonate, which helps stabilize the pH of the culture medium and prevents membrane structure from being oxidized and damaged, thus ensuring normal cell function. Example 3
[0019] In this embodiment, the basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth modifier including antibiotics 1%-3%; sustained-release agent including glycerol 6%-10%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%, the antioxidant including vitamin C, and the main directional differentiation inducer including dexamethasone.
[0020] Example 3, based on Example 2, increases the proportion of nitrogen source, growth factor, and slow-release agent while keeping the components unchanged. Through experiments, it was found that with increased nitrogen source and growth factor, the survival and reproduction rate of animal cells was greater than that of plant cells. At the same time, by increasing the proportion of slow-release agent, it was found that animal cells could still reproduce in an environment of 4°C, and the survival rate of animal cells was higher than that of animal cells in Example 2. Therefore, based on Example 2, Example 3 increases the proportion of nitrogen source, growth factor and slow-release agent while keeping the components unchanged, which can significantly improve the survival and growth rate of animal cells in an environment of 4°C.
[0021] Serum includes fetal bovine serum. Example 4
[0022] In this embodiment, the basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth regulators including antibiotics 3%-6%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and differentiation inducers 0.3%-0.8%, of which antioxidants include vitamin C and differentiation inducers include dexamethasone.
[0023] Example 4, based on Example 3, increased the proportion of antibiotic components while keeping the composition unchanged. Experiments showed that increasing the proportion of antibiotic components could effectively inhibit the proliferation and growth of animal cells, but simultaneously destroyed the activity of animal cells. Therefore, in Example 4, based on Example 3, the proportion of antibiotic components was increased while the composition remained unchanged. This enabled the antibiotic to significantly control the growth rate of animal cells at 4°C. Thus, it can be seen that the antibiotic does not impair the long-term preservation of the culture medium by the sustained-release agent at 4°C. Example 5
[0024] In this embodiment, the basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplement 5%-8%, basic fibroblast growth factor 6%-10%, and epidermal growth factor 4%-8%; growth modifier including antibiotics 1%-3%; sustained-release agent including glycerol 6%-10%; and special additives including antioxidants 5%-15% and differentiation inducers 0.3%-0.8%, where the antioxidants include vitamin C and the differentiation inducers include dexamethasone.
[0025] Example 5, based on Example 2, increased the proportions of growth factor supplement, basic fibroblast growth factor, epidermal growth factor, and slow-release agent while keeping the components unchanged. Experiments showed that with increased proportions of growth factor supplement, basic fibroblast growth factor, epidermal growth factor, and slow-release agent, the survival and reproduction rate of plant cells was greater than that of animal cells. Furthermore, increasing the proportion of slow-release agent demonstrated that plant cells could still reproduce at 4°C, and the survival rate of plant cells was higher than that of animal cells in Example 1. Therefore, based on Example 2, Example 3 increases the proportion of nitrogen source, growth factor and slow-release agent while keeping the components unchanged, which can significantly improve the survival and reproduction rate of plant cells in an environment of 4°C. Example 6
[0026] In this embodiment, the basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplement 5%-8%, basic fibroblast growth factor 6%-10%, and epidermal growth factor 4%-8%; growth regulators including antibiotics 3%-6%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and differentiation inducers 0.3%-0.8%, of which antioxidants include vitamin C and differentiation inducers include dexamethasone.
[0027] Example 6, based on Example 5, increased the proportion of antibiotic components while keeping the composition unchanged. Experiments showed that increasing the proportion of antibiotic components could effectively inhibit the reproduction and growth of plant cells, but at the same time destroyed the activity of plant cells. Therefore, in Example 6, based on Example 5, the proportion of antibiotic components was increased while the composition remained unchanged. This enabled the antibiotic to significantly control the growth rate of plant cells at 4°C. Thus, it can be seen that the antibiotic does not impair the long-term preservation of the culture medium by the sustained-release agent at 4°C.
[0028] In summary, the above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made within the scope of the claims of the present invention should be covered by the present invention.
Claims
1. A premixed liquid culture medium that can be stored at 4°C for a long period of time, comprising a basal culture medium and special additives, characterized in that: The basal culture medium includes carbon sources, nitrogen sources, inorganic salts, and growth factors; special additives include growth regulators and slow-release agents, as well as antioxidants and directional differentiation inducers.
2. The premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 1, characterized in that, The basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth modulators including antibiotics 1%-3%; sustained-release agents including glycerol 2%-8%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%.
3. The premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 1, characterized in that, The basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth modulators including antibiotics 1%-3%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%.
4. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 1, characterized in that, The basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplements 5%-8%, basic fibroblast growth factor 6%-10%, and epidermal growth factor 4%-8%; growth modulators including antibiotics 1%-3%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%.
5. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 3, characterized in that, The basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 2%-6%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 8%-12%, growth factor supplement 2%-6%, basic fibroblast growth factor 4%-7%, and epidermal growth factor 3%-6%; growth modulators including antibiotics 3%-6%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and differentiation inducers 0.3%-0.8%.
6. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 4, characterized in that, The basal culture medium and special additives include the following raw materials in parts by weight: carbon source including glucose 1.5%-2.5%, nitrogen source including peptone 0.8%-1.2%, inorganic salts including potassium dihydrogen phosphate 0.05%-0.15% and magnesium sulfate 0.02%-0.08%; growth factors including serum 3%-4%, growth factor supplements 5%-8%, basic fibroblast growth factor 6%-10%, and epidermal growth factor 4%-8%; growth modulators including antibiotics 3%-6%; sustained-release agents including glycerol 6%-10%; and special additives including antioxidants 5%-15% and directional differentiation inducers 0.3%-0.8%.
7. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to any one of claims 2 to 6, characterized in that: Antioxidants include vitamin C.
8. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to any one of claims 2 to 6, characterized in that: Dexamethasone is one of the agents that induce directional differentiation.
9. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to claim 3, characterized in that: Serum includes fetal bovine serum.
10. A premixed liquid culture medium that can be stored at 4°C for a long period of time according to any one of claims 2 to 6, characterized in that: Antioxidants also include sodium pyruvate.