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A serum-free medium and preparation method technology, applied in blood/immune system cells, vertebrate cells, animal cells, etc., can solve the problem of unstable medium components, and achieve the effect of good functional consistency and stable components
Inactive Publication Date: 2015-09-16
英普乐孚生物技术(上海)有限公司
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[0008] In view of the above-mentioned problems and / or other problems in the related art, the present invention provides a serum-free medium with more stable components and a preparation method thereof, which improves the instability of the medium components caused by the complex composition of albumin
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Embodiment 1～4
[0043] The preparation process of the serum-free culture medium of embodiment 1~4 is as follows:
[0044] Step 1: Obtain fatty acid-free albumin;
Embodiment 1
[0045]Embodiment 1 Get bovine serum albumin (the A1933 product purchased from SIGMA company) 1.5mmol, add water for injection and be made into 2000ml solution, add 50g gac, mix; Adopt the hydrochloric acid solution of 2mol / L to adjust the pH value to pH3.0; Place it in a water bath at 56°C for 30 minutes or place it overnight at 4°C; centrifuge (4000 rpm) to remove the activated carbon; then use 2mol / L NaOH solution to adjust the pH value to pH7.0; finally filter and sterilize to obtain the product of Example 1 fatty acid-free albumin solution;
Embodiment 2
[0046] Example 2 Take human serum albumin (purchased from Grifols (Grifols) company's human serum albumin (the product specification is 20% 50ml*10g / bottle, wherein the content of sodium octanoate is not more than 5mol / mol albumin)) 1.5 mmol, the specific operation process is the same as in Example 1, and the fatty acid-free albumin solution of Example 2 is obtained;
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Abstract
The invention relates to a serum-free medium and a preparation method thereof. The preparation method of the serum-free medium includes: loading fatty acid on FAF (fatty-acid-free) albumin, and then adding an obtained albumin solution into a basic culture medium. The serum-free medium obtained by using the preparation method of the serum-free medium is stable in component and good in functional consistency, and no difference can be caused due to difference of species origins or processing batches of the albumin.
Description
technical field [0001] The invention relates to a serum-free medium and a preparation method thereof, belonging to the technical field of cell culture. Background technique [0002] Serum-free medium refers to a synthetic medium that can maintain the growth and reproduction of cells in vitro for a long time without adding serum, and can be used to cultivate immune cells, such as lymphocytes (for example, T lymphocytes, B cells, NK cells, etc.) , dendritic cells (also known as DC cells), monocytes / macrophages, etc. [0003] Generally speaking, the nutrients required for the survival and expansion of immune cells in vitro include: albumin, transferrin, insulin, amino acids, vitamins, sugars, lipids, inorganic salts, growth factors, etc. Among these components, albumin has the greatest impact on cell growth and survival, and is also the component with the most complex function and composition. In serum-free medium, the function of albumin is mainly determined by the fatty aci...
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