Stem cell cryopreservation protective agent, preparation method and application
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A technology for cryopreservation and stem cells, applied in the medical field, can solve the problems of poor cryopreservation effect and high toxicity of cryopreservation, and achieve the effects of high possibility, high possibility of clinical application and convenient operation.
Pending Publication Date: 2020-08-28
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a cryoprotectant for stem cells, a prepa
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preparation example Construction
[0053] The preparation method of the stem cell cryoprotectant provided by one embodiment of the present invention comprises the following steps:
[0054] 1) adding trehalose to physiological saline or phosphate buffer to prepare a trehalose solution;
[0055] 2) Using glycerol as a solute and trehalose solution as a solvent to prepare a cryoprotectant for stem cells.
[0056] Based on the total amount of the cryoprotectant for stem cells, the concentration of trehalose in the cryoprotectant for stem cells is 0.25 mol / L-1.5 mol / L.
[0057] Further, step 1) also includes performing high-pressure steam sterilization on the obtained trehalose solution.
[0058] Physiological saline is a sodium chloride solution with a concentration of 0.9%.
[0059] Taking the total amount of phosphate buffer as the standard, the components of phosphate buffer are potassium dihydrogen phosphate 1.8mmol / L, disodium hydrogen phosphate 10mmol / L, sodium chloride 137mmol / L, and potassium chloride 2.7...
Embodiment 1
[0072] Prepare stem cell cryoprotectant according to the following formula:
[0073] Glycerol volume fraction is 20%;
[0074] Trehalose 1.0mol / L;
[0075] The solvent is saline.
Embodiment 2
[0077] Prepare stem cell cryoprotectant according to the following formula:
[0078] The volume fraction of glycerol is 10%;
[0079] Trehalose 0.3mol / L;
[0080] The solvent is saline.
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Abstract
The invention provides a stem cell cryopreservation protective agent which comprises the following components 10-30 vol% of glycerinum, 0.25-1.5 mol% of trehalose and a solvent being normal saline ora phosphate buffer solution. The stem cell cryopreservation protective agent provided by the invention is a cryopreservation protective agent with effective cryopreservation activity and function of stem cells, is efficient, non-toxic and free of xenogenic antigen, and has higher biological safety and clinical application possibility; no high-molecular polymer or biotoxic substance is used, and reagents used in the method have no cytotoxicity and are easy to elute; according to the invention, no blood product from any heterologous source exists, no heterologous antigen exists, and the possibility of hypersensitive reaction is low; compared with an existing cell cryopreservation protective agent, the cell cryopreservation protective agent is low in cost and large in large-dose application possibility; operation is convenient, and popularization is easy.
Description
technical field [0001] The invention relates to the medical field, in particular to a cryoprotectant for stem cells, a preparation method and an application. Background technique [0002] Stem cells have the potential to differentiate into mesoderm-derived cells such as adipogenic, osteogenic, and chondrogenic cells. At the same time, they can also secrete bioactive molecules such as growth factors and cytokines that stimulate tissue regeneration through paracrine signal transduction mechanisms. Alternative treatments for liver injury, refractory wounds, and autoimmune diseases, and even allogeneic transplantation, have become one of the most cutting-edge and hottest research fields in the medical field. If the stem cells can be preserved for a long time, the limitation of the timing of surgery can be well solved and a sufficient number of cells with stable genetic characteristics can be provided for future clinical applications. Cryopreservation is the main form of long-te...
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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 李青峰周双白张天钰
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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