Serum-free culture medium suitable for HEK293 cells
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A serum-free medium and medium technology, applied in the field of medium, can solve the problems of HEK293 cells that cannot be expanded at high density, continuous passage, affect cell viability, affect cell growth, etc., and achieve easy scale-up of culture and high protein production , reduce the effect of cell clumping
Pending Publication Date: 2021-10-22
武汉百齐生物技术有限公司
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[0006] In order to enable HEK293 cells to be used in transient transfection on a large scale, they are often domesticated to grow in suspension; however, during suspension culture, HEK293 cells are very easy to clump, and cell clumps directly affect cell viability, resulting in the inability of high-density expansion of HEK293 cells. serial passage
Glutamine in the existing anti-agglomeration HEK293 cell serum-free medium is easily degraded and affects cell growth. Therefore, it is urgent to develop a serum-free medium for HEK293 cells that is anti-agglomerative and does not affect cell growth.
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Embodiment 1
[0028] A serum-free medium suitable for HEK293 cells comprising amino acids, inorganic salts, vitamins, antisurbs and stabilizers. Each 1 L of medium contains 720 mg of amino acid, 25602.6 mg, vitamin 33.93 mg, additive 2615 mg, nucleoside 183 mg, F68600 mg, glutagro 200 mg, and the specific component and content are shown in Table 1.
[0029] The above components were subjected to aseptic deionized water, and the pH of the medium was adjusted with HCl and NaOH.
Embodiment 2
[0031] A serum-free medium suitable for HEK293 cells comprising amino acids, inorganic salts, vitamins, antisurbs and stabilizers.
[0032] Each 1 l was medium containing amino acid 1035 mg, inorganic salts 13310.9 mg, vitamin 62.5 mg, additive 1060 mg, nucleoside 587 mg, F68300 mg, Glutagro 400 mg, and water; specific components and content are shown in Table 1.
[0033] The above components were subjected to aseptic deionized water, and the pH of the medium was adjusted with HCl and NaOH to be 7.5.
Embodiment 3
[0035] A serum-free medium suitable for HEK293 cells comprising amino acids, inorganic salts, vitamins, antisurbs and stabilizers.
[0036] Each 1 l was medium containing amino acid 1235 mg, inorganic salts 18751.5 mg, vitamin 77.37 mg, additive 3250 mg, nucleoside 40 mg, F68800 mg, Glutagro 900 mg, and the remaining amount is water; specifically and the content is shown in Table 1.
[0037] The above components were subjected to sterile deionized water, and the pH of the medium was adjusted with HCl and NaOH.
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Abstract
The invention provides a serum-free culture medium suitable for HEK293 cells. The serum-free culture medium comprises amino acid, inorganic salt, vitamins, nucleoside, an additive, an anti-caking agent and a stabilizer; each 1 L of the culture medium contains 275 to 1650 mg of amino acid, 5470.2 to 37103 mg of inorganic salt, 10.92 to 124.4 mg of vitamin, 40 to 800 mg of nucleoside, 1060 to 3250 mg of an additive, 100 to 1000 mg of an anti-caking agent, 100 to 1000 mg of a stabilizer and the balance of water. The anti-caking agent is F68, and the stabilizing agent is glutaGRO. The culture medium disclosed by the invention is free of animal sources, does not contain proteins or growth factors, supports various HEK293 and derived cells thereof, has the effects of resisting caking, improving the cell stability and prolonging the shelf life, and can obtain higher protein yield in unit time.
Description
Technical field [0001] The present invention relates to the field of medium technologies, and more particularly to a serum-free medium suitable for use in HEK293 cells. Background technique [0002] A large number of high-quality recombinant proteins are required from various areas such as pharmaceutical and clinical applications from internally biochemical research. Techniques producing recombinant proteins with complex translation modifications represent the main challenges of structural and functional research. [0003] The rapid establishment of a mammalian cell line that can be used in transient transfection solves this obstacle. In the most commonly used mammalian cell line, human embryonic kidney 293 (HEK293) cells have become the standard choice for research and scale production due to the advantages of rapid growth, relatively easy to operate and high-transfection efficiency. [0004] Cell medium is the most important and direct environmental factor in determining the gr...
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