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Cryopreservation liquid for long-term preservation of human umbilical cord mesenchymal stem cells

A technology for long-term preservation of stem cells, which is applied in the field of cryopreservation for long-term preservation of human umbilical cord mesenchymal stem cells, which can solve the problems of unclear cell growth factors, pathogen cross-infection, cytotoxicity, etc., and improve safety and biological quality. Capacitive, reduced damage, less toxic effects

Pending Publication Date: 2021-05-28
朱灏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]The seed bank of human umbilical cord mesenchymal stem cells has been widely used clinically, but there is also the problem that it cannot be applied on a large scale due to the limitation of cryopreservation conditions. No exogenous contamination during storage is the key to the safety and reliability of human umbilical cord mesenchymal stem cells in clinical application. On the other hand, the clinical application of stem cells cultured with animal serum may cause cross-infection of pathogens. Therefore, it is necessary to develop a serum-free cryopreservation solution for human umbilical cord mesenchymal stem cells with clear components.
[0004]The long-term activity of human umbilical cord mesenchymal stem cell seed bank cells is closely related to the cryopreservation environment. Important impact, the existing cryopreservation solution has a greater toxic effect on cells

Method used

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  • Cryopreservation liquid for long-term preservation of human umbilical cord mesenchymal stem cells
  • Cryopreservation liquid for long-term preservation of human umbilical cord mesenchymal stem cells

Examples

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preparation example Construction

[0071] The second aspect of the present invention provides the preparation method of the cryopreservation solution for long-term preservation of human umbilical cord mesenchymal stem cells, comprising the following steps: adding pre-cooled compound electrolyte injection, chlorinated Sodium injection, trehalose solution, dextran, and vitamin C injection, mix well; then add pre-cooled human serum albumin, mix well, and get ready.

[0072] In a preferred embodiment, after the cryopreservation solution is prepared, a pre-cooling step is also included.

[0073] In a preferred embodiment, the pre-cooling is at 2-8°C for at least 30 minutes.

[0074] In a most preferred embodiment, the preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation solution comprises the following steps: slowly adding pre-cooled compound electrolyte injection, chlorinated Sodium injection, trehalose solution, dextran, vitamin C injection and heparan sulfate, mix well;...

Embodiment 1

[0080] Example 1 provides a cryopreservation solution for long-term preservation of human umbilical cord mesenchymal stem cells. The raw materials for the preparation of the cryopreservation solution, by volume percentage, at least include the following components: dimethyl sulfoxide 3%, 20% human serum albumin 20%, 0.9% sodium chloride injection 3%, 1mol / L trehalose solution 6%, dextran-40 glucose injection 24%, vitamin C injection 2%, human α1-acid Glycoprotein 25 μg / mL, heparan sulfate 25 μg / mL, compound electrolyte injection to make up the balance.

[0081] The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation solution comprises the following steps: slowly adding pre-cooled compound electrolyte injection, 0.9% sodium chloride injection, 1mol / L seaweed injection to dimethyl sulfoxide Sugar solution, dextran-40 glucose injection 24%, vitamin C injection and heparan sulfate, mix well; then slowly add pre-cooled human serum albumin...

Embodiment 2

[0083] Example 2 provides a cryopreservation solution for long-term preservation of human umbilical cord mesenchymal stem cells. The raw materials for the preparation of the cryopreservation solution, by volume percentage, at least include the following components: dimethyl sulfoxide 3%, 20% human serum albumin 10%, 0.9% sodium chloride injection 1%, 1mol / L trehalose solution 1%, dextran-40 glucose injection 5%, vitamin C injection 1%, human α1-acid Glycoprotein 20 μg / mL, heparan sulfate 400 μg / mL, compound electrolyte injection to make up the balance.

[0084] The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation solution comprises the following steps: slowly adding pre-cooled compound electrolyte injection, 0.9% sodium chloride injection, 1mol / L seaweed injection to dimethyl sulfoxide Sugar solution, dextran-40 glucose injection 24%, vitamin C injection and heparan sulfate, mix well; then slowly add pre-cooled human serum albumin...

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Abstract

The invention relates to the technical field of cryopreservation solutions, in particular to a cryopreservation solution for long-term preservation of human umbilical cord mesenchymal stem cells. The cryopreservation solution for long-term preservation of the human umbilical cord mesenchymal stem cells at least comprises the following preparation raw materials: dimethyl sulfoxide, human serum albumin, a compound electrolyte injection, a sodium chloride injection, a trehalose solution, dextran and a vitamin C injection. The cryopreservation liquid has small toxic and side effects on the human umbilical cord mesenchymal stem cells, can be directly used clinically, obviously improves the safety and biocompatibility of human umbilical cord mesenchymal stem cell retransfusion to human bodies, can improve the motility, proliferation and differentiation capabilities of the human umbilical cord mesenchymal stem cells on the premise of reducing the use amount of DMSO (dimethylsulfoxide), and can be directly retransfused after resuscitation, centrifugal washing is not needed, and damage to cells is reduced.

Description

technical field [0001] The invention relates to the technical field of cryopreservation solution, in particular to a cryopreservation solution for long-term preservation of human umbilical cord mesenchymal stem cells. Background technique [0002] Human umbilical cord mesenchymal stem cells refer to a kind of pluripotent stem cells isolated from umbilical cord. Compared with other sources of mesenchymal stem cells, they have obvious advantages in clinical application, such as easy to obtain materials, easy to collect, cryopreservation and relatively pure, etc. High differentiation potential, can be differentiated in multiple directions, such as osteoblasts, chondrocytes, hepatocytes, insulin-like cells, nerve cells and cardiomyocytes, etc., and there are no ethical issues. At present, human umbilical cord mesenchymal stem cells have been applied in the clinical field and achieved encouraging therapeutic effects. [0003] The seed bank of human umbilical cord mesenchymal ste...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 朱灏
Owner 朱灏
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