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Preparation method of human umbilical cord mesenchymal stem cell injection cryopreservation liquid

A technology of mesenchymal stem cells and cryopreservation solution, which is applied in the field of preparation of cryopreservation solution of human umbilical cord mesenchymal stem cell injection, can solve the problems of unclear cell growth factors, cross-infection of pathogens, and inability to apply on a large scale, so as to improve safety The effects of high biocompatibility, low toxicity and side effects, and optimization of the preparation method

Active Publication Date: 2021-06-04
朱灏
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  • Claims
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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, and at the same time has poor stability

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  • Preparation method of human umbilical cord mesenchymal stem cell injection cryopreservation liquid

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preparation example Construction

[0023] In order to solve the above technical problems, the first aspect of the present invention provides a method for preparing human umbilical cord mesenchymal stem cell injection cryopreservation liquid, which at least includes the following steps: adding glucose injection, chlorinated Sodium injection, dextran, and compound electrolyte injection were mixed uniformly; then human serum albumin was added and mixed uniformly to obtain a cryopreserved solution.

[0024] In a preferred embodiment, before the glucose injection, sodium chloride injection, dextran, compound electrolyte injection and human serum albumin are used for the preparation of the cryopreservation solution, pre-cooling treatment is also included.

[0025] In a preferred embodiment, after the cryopreservation solution is prepared, pre-cooling treatment is also included.

[0026] In a more preferred embodiment, the pre-cooling treatment is: storing at 1-10° C. for at least 30 minutes.

[0027] In a most prefe...

Embodiment 1

[0071] Embodiment 1 provides a human umbilical cord mesenchymal stem cell injection cryopreservation solution, the preparation raw materials of the cryopreservation solution, by volume percentage, at least include the following components: dimethyl sulfoxide 5%, compound electrolyte injection Liquid 56%, Dextran-40 Glucose Injection 8.5%, 0.9% Sodium Chloride Injection 3%, 10% Glucose Injection 2.5%, 20% Human Albumin 25%.

[0072] The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation liquid comprises the following steps: absorb DMSO and add it to a 500mL receiver, and slowly add pre-cooled 10% glucose injection, 0.9 sodium chloride injection, Dextran-40 Glucose Injection, Compound Electrolyte Injection, shake while adding until uniform; then slowly add pre-cooled 20% human serum albumin, mix upside down, pre-cooling to obtain; the pre-cooling is Pre-cool in a medical refrigerator at 4°C for 50 minutes.

Embodiment 2

[0074] Embodiment 2 provides a human umbilical cord mesenchymal stem cell injection cryopreservation solution, the preparation raw materials of the cryopreservation solution, by volume percentage, at least include the following components: dimethyl sulfoxide 5%, compound electrolyte injection Liquid 40%, Dextran-40 Glucose Injection 5%, 0.9% Sodium Chloride Injection 1%, 10% Glucose Injection 1%, 20% Human Albumin 48%.

[0075] The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation liquid comprises the following steps: absorb DMSO and add it to a 500mL receiver, and slowly add pre-cooled 10% glucose injection, 0.9 sodium chloride injection, Dextran-40 Glucose Injection, Compound Electrolyte Injection, shake while adding until uniform; then slowly add pre-cooled 20% human serum albumin, mix upside down, pre-cooling to obtain; the pre-cooling is Pre-cool in a medical refrigerator at 4°C for 50 minutes.

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Abstract

The invention relates to the technical field of cryopreservation liquid preparation, in particular to a preparation method of a human umbilical cord mesenchymal stem cell injection cryopreservation liquid. The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation liquid at least comprises the following steps: adding a glucose injection, a sodium chloride injection, dextran and a compound electrolyte injection into dimethyl sulfoxide, and uniformly mixing; and then adding the human serum albumin, and uniformly mixing to obtain the cryoprotectant. The preparation method of the human umbilical cord mesenchymal stem cell injection cryopreservation liquid is optimized, so that the prepared cryopreservation liquid has high stability, no floccules or precipitates are generated in the preparation process and after the preparation process is completed, the functional indexes and biological functions of human umbilical cord mesenchymal stem cells are maintained, and the safety and the biocompatibility of the human umbilical cord mesenchymal stem cell back transfusion to a human body are improved.

Description

technical field [0001] The invention relates to the technical field of cryopreservation solution preparation, in particular to a method for preparing a cryopreservation solution of human umbilical cord mesenchymal stem cell injection. 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 cor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 朱灏
Owner 朱灏
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