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Ultralow-temperature cryopreservation preservative for mesenchymal stem cells

A cryopreservation agent and a technology for stromal stem cells, which are applied in the field of ultra-low temperature cryopreservation agents for mesenchymal stem cells, can solve the problems of unsatisfactory stem cell cryopreservation solutions and different types of protective agents, and achieve excellent cell state and survival rate. improved effect

Inactive Publication Date: 2019-06-07
李冲杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, various universities, colleges, scientific research institutes, hospital platforms and other laboratories use different types of protective agents in the experimental process, and the ratio is also different. After freezing for different periods of time, the survival rate of cells during recovery is mostly 50%. %-80%, and the differentiation rate is mostly between 5%-15%. Combining the ratio of the two, it can be seen that the stem cell cryopreservation medium commonly used in various laboratories is not ideal

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  • Ultralow-temperature cryopreservation preservative for mesenchymal stem cells

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Experimental program
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Embodiment

[0020] 1. Main instruments, reagents and consumables

[0021] Instruments: Thermo CO2 incubator, Thermo centrifuge, SD-WJ-2HD ultra-clean workbench, Accuri C6plus flow cytometer, JSY-SC-021H cell counter, East Asia liquid nitrogen tank.

[0022] Reagents: GENVIEW PBS, Gibco pancreatin, Gibco penicillin / streptomycin, Helios serum, Gibco MEM medium, heparin sodium, DMSO, 12% HES water solution, 20% ALB solution, mass volume fraction It is a 16% trehalose aqueous solution, ethylene glycol.

[0023] Consumables: T175 cell culture flask, 10ml-50ml pipette, 50ml centrifuge tube, cryotube.

[0024] 2. Method steps

[0025] (1) Subculture UCMSC in the laboratory to the P4 generation, digest and collect for cryopreservation, and take 10ul and 100ul for cell count and flow cytometry respectively.

[0026] (2) Prepare the following freezing solutions:

[0027] Group A: 10ml DMSO, 90ml Gibco MEM medium;

[0028] Group B: 10ml DMSO, 10ml 20% ALB solution, 80ml Gibco MEM medium;

[0029] Group C: 10ml o...

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Abstract

The invention discloses an ultralow-temperature cryopreservation preservative for mesenchymal stem cells. The ultralow-temperature cryopreservation preservative is characterized by comprising the following ingredients in parts by volume: 8-10 parts of DMSO and 5-6 parts of 12%-15% HES aqueous solution. The ultralow-temperature cryopreservation preservative has the beneficial effects that the survival rate of the stem cells cryopreserved by the ultralow-temperature cryopreservation preservative is increased remarkably after resuscitation after cryopreservation of a certain cycle; and a cell state of the stem cells cryopreserved by the ultralow-temperature cryopreservation preservative is excellent after resuscitation after cryopreservation of a certain cycle.

Description

Technical field [0001] The invention relates to the field of cryopreservation protective agents, in particular to a cryopreservation protective agent for mesenchymal stem cells. Background technique [0002] Umbilical Cord Mesenchymal Stem Cells (UCMSCs) refer to a kind of multifunctional stem cells derived from neonatal umbilical cord tissue. UCMSC can differentiate in multiple directions, in bone, cartilage, muscle, tendon, ligament, Nerve, liver, endothelium and myocardium have broad clinical application prospects. [0003] Freezing cells, tissues, etc. in an environment below -80°C causes the cell's biochemical process and enzyme metabolism to almost completely stop to ensure the cell's viability and time. This process is called cell cryopreservation. UCMSC often needs long-term cryopreservation in the process of experiment and production. The biggest threat of cryopreservation to cells is the damage to cell membranes and organelles caused by water crystals at low temperatures...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 李冲杰孙洪霞李宾宾侯文文
Owner 李冲杰