Serum-free medium and preparation method thereof

A serum-free medium and amino acid technology, applied in the field of biochemistry, can solve the problems of high price and achieve the effects of fast adaptability, fast wall attachment and simplified extraction process

Inactive Publication Date: 2017-05-31
山东巨山能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researchers to study how different types of blood clot (blood vessels) form when they donate their body fluids into specialized tissue called stem or bone marrow stromal vascular fraction(BSF). These experiments help scientists better understanding disease processes related to these materials by studying them under specific conditions.

Problems solved by technology

This patented technical problem addressed in this patent relates to developing an economically viable solution that allows for efficient use of animal blood plasma (BP) without adding any unnecessary materials or steps during upkeeping processes.

Method used

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  • Serum-free medium and preparation method thereof
  • Serum-free medium and preparation method thereof
  • Serum-free medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The present invention prepares the step of serum-free culture medium as follows:

[0042] (1) Preparation of stock solution:

[0043] Glycine, L-alanine, L-arginine hydrochloride, L-asparagine monohydrate, L-aspartic acid, L-cysteine ​​hydrochloride monohydrate, L-cystine Dihydrochloride, L-Glutamic Acid, L-Histidine Hydrochloride Monohydrate, L-Isoleucine, L-Leucine, L-Lysine Hydrochloride, L-Methionine , L-phenylalanine, L-proline, L-serine, L-threonine and L-valine are dissolved in water respectively or two or more are mixed and dissolved in water to prepare amino acid stock solution, and carry out cryopreservation;

[0044] Dissolving L-glutamine in water to prepare a glutamine stock solution and storing it in a frozen state;

[0045] Dissolving L-tryptophan and L-tyrosine disodium salt separately or mixed in 0.1mol / L hydrochloric acid to prepare tryptophan and tyrosine disodium salt stock solution, and cryopreservation;

[0046] Dissolve ascorbic acid, biotin, ...

Embodiment 2

[0059] The difference between this embodiment and Embodiment 1 is that the concentration (mg / L) of the above-mentioned raw materials in the respective stock solutions in step (1) is as follows:

[0060]

[0061]

[0062] The temperature for refrigerated storage is 6°C;

[0063] The concentration (mg / L) of each raw material in the serum-free medium in the step (2) is as follows:

[0064]

[0065]

Embodiment 3

[0067] The difference between this embodiment and Embodiment 1 is that the concentration (mg / L) of the above-mentioned raw materials in the respective stock solutions in step (1) is as follows:

[0068]

[0069]

[0070] The temperature for refrigerated storage is 8°C;

[0071] The concentration (mg / L) of each raw material in the serum-free medium in the step (2) is as follows:

[0072]

[0073]

[0074] Taking Vero cells as an example, a comparative test was carried out. The test was carried out in an environment with a temperature of 37° C. for 3 days, and the influx of CO2 was 5.0%. Wherein, the medium used in Comparative Example 1 is MEM medium+10% calf serum, and the medium used in Comparative Example 2 is F12 medium+10% calf serum. The experimental data are as follows:

[0075]

[0076] From the above test data, it can be seen that the main performance of the serum-free medium of the present invention in terms of cell cultivation is not inferior to that o...

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Abstract

The invention discloses a serum-free medium and a preparation method thereof, and belongs to the technical field of biochemistry. The preparation method comprises the following steps: 1, preparing an amino acid stock solution, a glutamine stock solution, a tryptophan and tyrosine disodium salt stock solution, a vitamin stock solution, an inorganic salt stock solution, a trace element stock solution and an insulin stock solution; and 2, adding a proper amount of potassium chloride, sodium chloride, and the amino acid stock solution, the tryptophan and tyrosine disodium salt stock solution, the vitamin stock solution, the inorganic salt stock solution, the trace element stock solution and the insulin stock solution which are prepared in step 1 into ultrafine water, adding sodium bicarbonate, carrying out mixing and dissolving to obtain a mixed solution, and adding the glutamine stock solution which is unfrozen by 37 DEG C water bath into the mixed solution before use in order to prepare the serum-free medium. The serum-free medium makes cell proliferation normal and the activity stable, is non-obviously different from a medium containing 10% of a finished calf serum product, has an obviously lower protein content than routine media, simplifies the extraction technology, and reduces the production cost.

Description

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Claims

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Application Information

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Owner 山东巨山能源科技有限公司
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