Cell culture medium and cell culture method
A cell culture and culture medium technology, applied in the field of cell culture medium and cell culture, can solve the problems of low fibroblast survival rate, inconvenient laboratory operation, affecting the growth of fibroblasts, preservation of proliferation efficiency, etc.
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Embodiment 1
[0048] This embodiment provides a cell culture medium, comprising: culture medium, keratin particles and serum, wherein the serum is bovine serum or autologous serum.
[0049] Wherein, the nutrient solution is preferably DMEM (Dulbecco's Modified Eagle Medium) nutrient solution, and the nutrient solution is added to the cell culture medium described in this embodiment, which is rich in nutrients and can provide cells with the nutrients needed for growth, providing cells with Provide a suitable growth environment.
[0050] Specifically, the culture fluid preferably includes the following components by mass: 90-110 parts of DMEM dry powder, 30-50 parts of sodium bicarbonate, 1-5 parts of penicillin, 1-5 parts of streptomycin, 30-50 parts of HEPES and L-glutamine 1-5 parts.
[0051] In practical applications, the mass parts of DMEM dry powder can be 90 parts, 95 parts, 100 parts, 105 parts, 110 parts, etc., preferably 100 parts; the mass parts of sodium bicarbonate can be 30 par...
Embodiment 2
[0065] On the basis of Example 1, this example provides a cell culture medium, which further includes chitosan and / or collagen.
[0066] Chitosan is a basic polysaccharide with biodegradability, cell affinity and biological effect, and contains free amino groups. Chitosan is easy to dissolve in culture medium and serum, and the dissolved solution also contains amino (NH 2 + ), these amino groups can inhibit the activity of bacteria by combining negative electrons and maintain a safe environment for cell growth. In this embodiment, the concentration of chitosan is 0-1%, specifically, it can be 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0% % etc., depending on specific circumstances, the present application does not limit this, preferably 0.3%.
[0067] Collagen is the main component of biopolymers and animal connective tissue, and it is also the most abundant and widely distributed functional protein in mammals. It has good biocompatibility, biodegradability and...
Embodiment 3
[0070] Such as figure 1 As shown, this embodiment provides a cell culture method, including step S1 to step S4.
[0071] S1. Take out the skin tissue block, rinse the skin tissue block, transfer it to a petri dish, add digestive juice, and leave overnight.
[0072] Specifically, the skin tissue block can be washed 3 times to remove fat and blood, transferred to a petri dish, and added with 0.25% neutral protease digestion solution overnight at 4°C.
[0073] Wherein, the 0.25% neutral protease digestion solution (Dispase II) is obtained by dissolving neutral protease into phosphate buffered saline solution and adjusting the pH to 7.4.
[0074] In practical applications, the pH of the phosphate buffered saline (PBS) is preferably 7.2, and the phosphate buffered saline may include sodium chloride, potassium chloride, disodium hydrogen phosphate dodecahydrate and potassium hydrogen phosphate.
[0075] In this embodiment, the neutral protease digestion solution can improve the de...
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Abstract
Description
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Application Information
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