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Mesenchymal stem cell resuscitation protective solution, preparation method and resuscitation method

A technology of stem cells and protective solution, applied in the field of cells, can solve the problems of low cell recovery rate and biological activity, and achieve the effects of improving biological activity and recovery rate, increasing adherence rate and reducing damage

Inactive Publication Date: 2019-03-19
CHENGDU QINGKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the above-mentioned problems existing in the prior art, the present invention provides a mesenchymal stem cell resuscitation protection solution, a preparation method and a resuscitation method, which can effectively solve the problems of low cell recovery rate and biological activity of mesenchymal stem cells after resuscitation

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  • Mesenchymal stem cell resuscitation protective solution, preparation method and resuscitation method

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Embodiment 1

[0025] A mesenchymal stem cell recovery protection solution, comprising cell culture stock solution, fresh medium, L-carnitine and trehalose, the pH value of the recovery protection solution is 7.2-7.6; wherein, the volume ratio of fresh medium and cell culture stock solution is 1:1, the final concentrations of L-carnitine and trehalose were 1 mg / mL and 0.2 mg / mL, respectively.

[0026] The above-mentioned fresh medium is a fresh complete medium (manufactured by Gibco); the cell culture stock solution is cultured placental mesenchymal stem cells, and the remaining medium after the cells are separated (the separated cells are used for cryopreservation, and used for recovery) The cell culture stock solution in the resuscitation protection solution is the cell stock solution that has cultivated this batch of cells), the storage period of the cell culture stock solution is no more than 1 year, and the content of VEGF (vascular endothelial growth factor) is 286.09pg / mL, EGF (epiderm...

Embodiment 2

[0035] A mesenchymal stem cell recovery protection solution, comprising cell culture stock solution, fresh medium, L-carnitine and trehalose, the pH value of the recovery protection solution is 7.2-7.6; wherein, the volume ratio of fresh medium and cell culture stock solution is 1:1, the final concentrations of L-carnitine and trehalose were 3mg / mL and 0.2mg / mL, respectively.

[0036] The above-mentioned fresh medium is a fresh complete medium (manufactured by Gibco); the cell culture stock solution is cultured placental mesenchymal stem cells, and the remaining medium after the cells are separated (the separated cells are used for cryopreservation, and used for recovery) The cell culture stock solution in the resuscitation protection solution is the cell stock solution that has cultivated this batch of cells), the storage period of the cell culture stock solution is no more than 1 year, and the content of VEGF (vascular endothelial growth factor) is higher than 260pg / mL, EGF (...

Embodiment 3

[0045] A mesenchymal stem cell recovery protection solution, comprising cell culture stock solution, fresh medium, L-carnitine and trehalose, the pH value of the recovery protection solution is 7.2-7.6; wherein, the volume ratio of fresh medium and cell culture stock solution is 1:1, the final concentrations of L-carnitine and trehalose were 5mg / mL and 0.2mg / mL, respectively.

[0046] The above-mentioned fresh medium is a fresh complete medium (manufactured by Gibco); the cell culture stock solution is cultured placental mesenchymal stem cells, and the remaining medium after the cells are separated (the separated cells are used for cryopreservation, and used for recovery) The cell culture stock solution in the resuscitation protection solution is the cell stock solution that has cultivated this batch of cells), the storage period of the cell culture stock solution is no more than 1 year, and the content of VEGF (vascular endothelial growth factor) is higher than 260pg / mL, EGF (...

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Abstract

The invention provides a mesenchymal stem cell resuscitation protective solution, a preparation method and resuscitation method. The resuscitation protective solution is prepared from a cell culture stock solution, a fresh culture medium, L-carnitine and mycose. The resuscitation method comprises the following steps: taking out a cryopreserved mesenchymal stem cell, placing at the temperature of 42-45 DEG C until the mesenchymal stem cell starts to thaw; adding the resuscitation protective solution, uniformly mixing, centrifuging, and washing precipitate; adding the resuscitation protective solution into the washed mesenchymal stem cell, regulating the cell density, culturing, replacing a complete culture medium after 24 hours, and continuing culture by a conventional technology. Accordingto the resuscitation method, the resuscitation protective solution consisting of specific ingredients can be used for resuscitating cells and used as a cell washing solution due to the synergistic effect of the ingredients. The mesenchymal stem cell is resuscitated by combining the resuscitation protective solution and the resuscitation method, the cell reclamation rate and biological activity after cell resuscitation can be remarkably improved.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a mesenchymal stem cell resuscitation protection solution, a preparation method and a resuscitation method. Background technique [0002] Mesenchymal stem cells are stem cells with self-replication ability and multi-lineage differentiation ability. They have the advantages of low immunogenicity and strong cell primitiveness, and have become important seed cells in cell therapy. Cryopreservation and recovery are the basis for mesenchymal stem cells to play a therapeutic role, especially the recovery rate and biological activity of cells after recovery have an important impact on the therapeutic ability of mesenchymal stem cells. At present, the resuscitation of mesenchymal stem cells generally uses physiological saline, PBS buffer or medium for washing, which has the disadvantages of low cell recovery rate and biological activity after resuscitation. Therefore, it is an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/073C12N5/0775
CPCC12N5/0605C12N5/0667C12N5/0668C12N5/0682C12N2500/34C12N2501/105C12N2501/11C12N2501/115C12N2501/12C12N2501/148C12N2501/15C12N2501/165C12N2501/999
Inventor 高雪华张学超海泉
Owner CHENGDU QINGKE BIOTECH
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