Proliferation culture medium for human skin fibroblasts and preparation method thereof
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A technology for fibroblasts and culture medium, applied in the field of culture medium, can solve the problems of serum safety hazards, complex serum components, low cell proliferation rate, etc., achieves simple and easy operation in the preparation process, improves cell growth rate, and improves proliferation efficiency. Effect
Inactive Publication Date: 2021-02-23
郑州佐爵生物科技有限公司
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[0005] The traditional culture of skin fibroblasts is to add serum to the basal medium, but the use of serum will bring risks and hidden dangers: the serum contains cytostatic factors and toxic factors, which have potential cytotoxicity; in addition, the serum components are complex, There are differences in quality between batches, which increases the difficulty and instability of product quality control; potential pathogenic microorganisms in serum (such as prions that cause bovine spongiform encephalitis) bring safety hazards that cannot be ignored to the use of serum In addition, the protein in the serum medium is difficult to be completely removed by ordinary separation methods, resulting in high difficulty in the separation and purification of cell expression products, which seriously affects the final quality of the product
These unfavorable factors have prompted the improvement of the culture method, and the serum-free medium is used to replace the serum medium. The existing serum-free medium has problems such as complex components, high application cost, poor stability, and low cell proliferation rate. Improve
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Embodiment 1
[0021] An expansion medium for human skin fibroblasts, consisting of DMEM / F12 medium and additives, the concentration of the additives in the DMEM / F12 medium is: insulin 45 μg / mL, phytic acid 18 μg / mL, Germinated brown rice powder 20 μg / mL, feruloylglycan 15 ng / mL, nano zinc oxide with an average particle size of 20 nm ng / mL, chitin 15 μg / mL, sodium selenite 18 μg / mL.
[0022] The preparation method of the above-mentioned expansion medium for human skin fibroblasts comprises the following steps: taking the basal medium and adding insulin, phytic acid, germinated brown rice flour, feruloylglycan, nano-zinc oxide, chitin, and selenous Sodium acid, mix evenly, and use 0.22μm microporous membrane to filter and sterilize.
Embodiment 2
[0024] An expansion medium for human skin fibroblasts, consisting of DMEM / F12 medium and additives, the concentration of the additives in the DMEM / F12 medium is: insulin 50 μg / mL, phytic acid 20 μg / mL, Germinated brown rice flour 25 μg / mL, feruloylglycan 18 ng / mL, nano zinc oxide with an average particle size of 20 nm 8 ng / mL, chitin 17 μg / mL, sodium selenite 20 μg / mL.
[0025] The preparation method of the expansion medium for human skin fibroblasts is the same as in Example 1.
Embodiment 3
[0027] An expansion medium for human skin fibroblasts, consisting of DMEM / F12 medium and additives, the concentration of the additives in the DMEM / F12 medium is: insulin 55 μg / mL, phytic acid 25 μg / mL, Germinated brown rice flour 27μg / mL, feruloylglycan 20ng / mL, nano zinc oxide with an average particle size of 50nm 10ng / mL, chitin 20μg / mL, sodium selenite 21μg / mL.
[0028] The preparation method of the expansion medium for human skin fibroblasts is the same as in Example 1.
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Abstract
The invention discloses a proliferation culture medium for human skin fibroblasts. The proliferation culture medium comprises a basal culture medium and an additive. The additive comprises the following raw materials of insulin, inositol hexaphosphate, germinated brown rice powder, feruloyl glycan, nano-zinc oxide, chitin and sodium selenite. The proliferation culture medium for the human skin fibroblasts is clear in components, free of exogenous serum and animal-derived protein and higher in safety. The inositol hexaphosphate and germinated brown rice powder in the additive are beneficial tomaintaining the proliferation stability of the human skin fibroblasts in the proliferation process and increasing the cell growth rate. The feruloyl glycan and nano-zinc oxide in the additive can improve the proliferation activity of the human skin fibroblasts and avoid cell apoptosis, so that the proliferation efficiency is improved. The chitin and sodium selenite are also added, so that an antioxidant protection effect on cells is achieved. The invention further provides a preparation method of the proliferation culture medium for the human skin fibroblasts. The preparation process is simpleand convenient, and commercialization is facilitated.
Description
technical field [0001] The invention relates to a medium, in particular to an expansion medium for human skin fibroblasts and a preparation method thereof. Background technique [0002] Fibroblasts are the main cellular components of loose connective tissue, differentiated from embryonic mesenchymal cells. Fibroblasts have strong functional activities, and have obvious protein synthesis and secretion activities. Under certain conditions, they can achieve mutual transformation with fibroblasts. Fibroblasts play an important role in the repair of different degrees of cell degeneration, necrosis, tissue defect and bone trauma. The unique properties of fibroblasts help repair damaged skin and reduce the effects of aging on the skin. [0003] Skin fibroblasts are widely distributed in the dermis. Fibroblasts secrete a variety of substances to form the extracellular matrix, thereby forming connective tissue and exerting structural functions on the body. Fibroblasts play an impo...
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