General serum-free crypreservation solution for MSCs (mesenchymal stem cells) and preparation method of crypreservation solution
A technique of mesenchymal stem cells and cryopreservation method is applied in the field of universal serum-free cryopreservation of mesenchymal stem cells and its preparation, which can solve problems such as low toxicity, and achieve the effect of avoiding complement reaction and avoiding potential risks of virus infection.
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[0046] The invention also discloses a method for preparing a universal serum-free cryopreservation solution for mesenchymal stem cells, comprising the following steps:
[0047] Step 1. Weighing: Weigh the following components according to volume percentage: compound dextran 40 injection 0.5-10%, hydroxyethyl starch 130 / 0.41%-10%, glycerin 5%-30%, serum-free medium 25% -70%, serum replacement 0.5%-1%, human albumin 20%-40%, vitamin C 0.05%-1%, vitamin E 0.2%-1%, non-essential amino acid 0.5%-3%, the above quality The total percentage content is 100%;
[0048] Step 2. In a biosafety cabinet that is working normally (the air purification LED light turns green), prepare a sterile pyrogen-free serum square bottle, add each component in turn and mix well;
[0049] Step 3. Use a 0.2um filter to aseptically filter the mixed solution above into a new sterile pyrogen-free serum square bottle, and take samples for microbial testing, endotoxin testing, and mycoplasma testing. is qualifi...
Embodiment 1
[0062] A universal serum-free cryopreservation solution for mesenchymal stem cells, which consists of the following components according to volume percentage: 0.5% compound dextran 40 injection, 1.5% hydroxyethyl starch (130 / 0.4), 5% glycerol, and serum-free medium 70%, serum substitute 0.5%, human serum albumin 20%, vitamin C 1%, vitamin E 1%, non-essential amino acid 0.5%, the total amount of the above mass percentages is 100%.
[0063] Among them, the serum-free medium is Lonza UItraCULTURE TM ; Serum replacement is Pall Ultroser TM .
[0064] The above-mentioned universal serum-free cryopreservation solution for mesenchymal stem cells is prepared by the following method:
[0065] Step 1, weighing: Weigh each component according to the above-mentioned volume percentage;
[0066] Step 2. Prepare a sterile pyrogen-free serum square bottle, add each component in sequence and mix well;
[0067] Step 3. Use a 0.2um filter to aseptically filter the mixed solution above into a...
Embodiment 2
[0082] A universal serum-free cryopreservation solution for mesenchymal stem cells, which consists of the following components according to volume percentage: compound dextran 40 injection 10%, hydroxyethyl starch (130 / 0.4) 10%, glycerol 30%, serum-free medium 25%, serum substitute 1%, human serum albumin 23.25%, vitamin C 0.05%, vitamin E 0.2%, non-essential amino acid 0.5%, the total content of the above mass percentages is 100%.
[0083] Among them, the serum-free medium is Lonza UItraCULTURE TM ; Serum replacement is Pall Ultroser TM .
[0084] The above-mentioned universal serum-free cryopreservation solution for mesenchymal stem cells is prepared by the following method:
[0085] Step 1, weighing: Weigh each component according to the above-mentioned volume percentage;
[0086] Step 2. Prepare a sterile pyrogen-free serum square bottle, add each component in sequence and mix well;
[0087] Step 3. Use a 0.2um filter to aseptically filter the mixed solution above into...
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