How cell media are used.

There are many kinds of cell culture, cell culture mode is also more, how to use the culture base correctly and maximize the nutrition of the culture to maintain the growth of cells, is very important for each culturer.
use of culture media varies according to different media types, culture methods and cell types.
, the composition of cell culture fluid cell culture fluid refers to the liquid environment of cell growth, generally refers to the complete culture fluid, added serum, hydrolytic emulsion, various additives, etc. can be used directly cultured liquid.
1. Cell media and serum pattern serums are needed for most cell line that grow and multiply in traditional media formulations, because most individual media do not provide all the nutrients needed for cell growth, such as MEM media, the more commonly used amount is 5% to 10%, and the amount of special low serum medium serum can be reduced to about 3%, cell form and function are not affected.
2, cell culture medium and emulsion model chemical synthesis medium is now used on a large scale, but in some culture areas hydrolytic emulsion is still used to supplement the lack of amino acids, small peptide substances in the culture.
The more common way to do this is to dissolve the hydrolyzed milk protein (generally at a concentration of 5 inches) using Hanks' BSS or Earle's BSS balance, i.e. to form a lactobacid (European) liquid, which is then used proportionally with a chemically synthesized culturebase (generally MEM).
3, cell media and various additives (growth factors, etc.) model composition complex media also contain many compounds, including proteins, peptides, nucleotides, citric acid cycle intermediates, acetone acid and lipids.
have found that these ingredients are necessary when serum concentrations in the culture are reduced.
in the case of serum, these components can also help cell cloning culture and maintain the growth of certain specific cell linens.
hormones and growth factors are not specified in most conventional medium formulations, but they are usually added in serum-free mediums to replace hormones in serums that increase the rate of adage of many different types of cells; Family, EGF, PDGF, IGF-1 and IGF-2 and interleukin, growth factors and cytokines have a wide range of specificity, generally with hormones or other substances to synergy or addition.
Another common addition is serum substitutes, there are many commercial products that can completely or partially replace the serum ingredients in traditional culture fluids, its stability is better, but there are still differences between batches, the composition of the product is not very certain.
2, cell culture base preparation 1, dry powder culture base primary peptide preparation 1) preparation filter sterilization cell culture base (1) read the instructions for the use of the culture bed to determine what additives need to be added (e.g. NaHCO3, L-glutamine, sodium acetone, HEPES, etc.).
(2) Pour all the medium into a container as needed, wash off the residual medium in the bag with a small amount of injectable water (20 to 30 degrees C) and put it into the container.
water for injection to 95% of the total volume, stirring slightly to dissolve.
(3) add the specified amount of sodium bicarbonate and the substance to be added.
(4) stir to dissolve, add water injection to the specified volume.
(5) pH to the desired value with 1 mol / L sodium hydroxide solution or 1 mol / L hydrochloric acid solution.
(6) the sterilization is filtered with a positive pressure of 0.22 ?m membrane.
(7) the solution should be preserved in light at 2 to 8 degrees C.
specific method of use refers to the instructions on the base bag.
2) Preparation of autoclactive cell medium (1) According to the need to pour all the medium into a container, with a small amount of injection water (20 degrees C to 30 degrees C) to wash the residual medium in the bag, into the container.
water for injection to 95% of the total volume, stirring slightly to dissolve.
(2) sterilized for 15 minutes at 121 degrees C and 15psi.
(3) To be cooled to room temperature, add sterile 0.2 mol / L L-glutamine solution specified volume, sterile 7.5% (w/v) sodium bicarbonate solution specified volume, plus injection water (20 degrees C to 30 degrees C) to the specified volume, mixed.
(4) If necessary, adjust pH to the desired value with 1 mol / L sodium hydroxide solution or 1 mol / L hydrochloric acid solution.
(5) the solution should be preserved in light at 2 to 8 degrees C.
specific method of use refers to the instructions on the base bag.
2, Note 1) Cell culture water is generally three steamed water (distilled by quartz distiller) or ultra-pure water, pharmaceutical production is generally for injection water.
2) It is recommended to use packaging that matches the amount used as much as possible, as once the package is opened, the parts may become quality or group due to moisture.
3) When dissolving the dry powder culture, add 90%-95% of the final volume of culture water, additives added and then replenished.
4) If you need to add more parts, one group dissolves completely and then adds another.
5) Add sodium bicarbonate until the base is completely dissolved to avoid precipitation.
6) Once all ingredients have been completely dissolved, the methyl should be filtered or high-pressure sterilized immediately to avoid precipitation or microbial contamination.
7) When filtering sterilization, pH should generally be adjusted to 0.1 to 0.2 units lower than the desired value, because after filtration sterilization, the pH will increase by about 0.2.
8) During cell culture, it is recommended to reduce the amount of antibiotics added without or without a small amount of antibiotics, such as lower concentrations of serum.
9) It is recommended to use 1 N HCl or 1 N NaOH to regulate the pH of the media, as sodium bicarbonate is used to regulate the osmosis pressure of the culture fluid.
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