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.

Inactive Publication Date: 2018-12-28
成都汇欣生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, the present invention provides a general-purpose serum-free cryopreservation solution for mesenchymal stem cells and a preparation method thereof, which avoid Potential risks from low toxicity and complement response of some components of existing generic methods

Method used

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  • General serum-free crypreservation solution for MSCs (mesenchymal stem cells) and preparation method of crypreservation solution
  • General serum-free crypreservation solution for MSCs (mesenchymal stem cells) and preparation method of crypreservation solution
  • General serum-free crypreservation solution for MSCs (mesenchymal stem cells) and preparation method of crypreservation solution

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preparation example Construction

[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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Abstract

The invention discloses a general serum-free crypreservation solution for MSCs (mesenchymal stem cells) and a preparation method of the crypreservation solution. The crypreservation solution is prepared from components in percentage by volume as follows: 0.5%-10% of a compound dextran 40 injection , 1%-10% of hydroxyethyl starch (130 / 0.4), 5%-30% of glycerin, 25%-70% of a serum-free medium, 0.5%-1% of a serum substitute, 20%-40% of human albumin, 0.05%-1% of vitamin C, 0.2%-1% of vitamin E and 0.5%-3% of nonessential amino acid. By the aid of the crypreservation solution, on the premise that the cell activity of MSCs under the deep hypothermia cryopreservation condition is not lower than that in existing methods, potential risks caused by low toxicity and complement reactions of certain components in existing general methods are avoided.

Description

technical field [0001] The invention belongs to the field of regenerative medicine biotechnology, and in particular relates to a universal serum-free cryopreservation solution for mesenchymal stem cells and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSC, mesenchymal stem cells) are an important member of the stem cell family, derived from the mesoderm in the early development stage, and belong to pluripotent stem cells. MSC was originally found in the bone marrow because of its multi-directional differentiation potential, hematopoietic support and promotion Stem cell implantation, immune regulation and self-replication have attracted increasing attention. [0003] For example, mesenchymal stem cells can be differentiated into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells under specific induction conditions in vivo or in vitro, after continuous subculture and cryopreser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 谭汝福
Owner 成都汇欣生命科技有限公司
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