Umbilical cord mesenchymal stem cell protein-free non-programmed freezing medium and preparation method thereof
A high-quality stem cell, protein-free technology, applied in the field of stem cell culture, can solve the problems of decreased cell expansion ability, time-consuming and laborious, high cost, etc., to improve the recovery rate and vitality, avoid the body's immune response, and reduce mutual extrusion Effect
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Embodiment 1
[0034] Preparation method of 1L of umbilical cord mesenchymal stem cell protein-free non-programmed cryopreservation solution:
[0035] 1. Preparing the base solution: Prepare the base solution according to the DMEM / F-12 formula, in which the glucose content is reduced to 1000mg / L, and the contents of other components remain unchanged.
[0036] 2. Weigh 0.4mg sodium molybdate dihydrate, 0.1mg nickel chloride, 0.15mg manganese dichloride tetrahydrate, 0.28mg cholesterol, 0.36mg linoleic acid, 6mg linolenic acid, 2.5mg spermine hydrochloride, 4g Progesterone, 10g erythritol, 15g trehalose, 25nmol Q-VD-OPh, 7.5mg sodium selenite, 0.03mg verbisoflavone were dissolved in the base solution in turn;
[0037] 3. Add 50ml of dimethyl sulfoxide, 10mg of sodium borate hydrate, 50g of hydroxyethyl starch, 40g of polyvinylpyrrolidone, 10g of dextran, and 15g of Lutrol F68 to it in turn, and fully dissolve;
[0038] 4. Adjust the pH to 6.9 with hydrochloric acid, then dilute the volume to ...
Embodiment 2
[0040] Preparation method of 1L of umbilical cord mesenchymal stem cell protein-free non-programmed cryopreservation solution:
[0041] 1. Preparing the base solution: Prepare the base solution according to the DMEM / F-12 formula, in which the glucose content is reduced to 1000mg / L, and the contents of other components remain unchanged.
[0042] 2. Weigh 0.8mg sodium molybdate dihydrate, 0.2mg nickel chloride, 0.05mg manganese dichloride tetrahydrate, 0.1mg cholesterol, 0.1mg linoleic acid, 0.5mg linolenic acid, 4mg spermine hydrochloride, 1.5mg g progesterone, 6 g erythritol, 20 g trehalose, 30 nmol Q-VD-OPh, 5 mg sodium selenite, 0.02 mg mullein isoflavones were dissolved in the base solution in turn;
[0043] 3. Add 45ml of dimethyl sulfoxide, 40mg of hydrated sodium borate, 30g of hydroxyethyl starch, 30g of polyvinylpyrrolidone, 20g of dextran, and 30g of Lutrol F68 successively to it, and fully dissolve;
[0044] 4. Adjust the pH to 6.9 with hydrochloric acid, then dilut...
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