cells temporarily leave the growth state and its cell characteristics to survive, so that the necessary time to wake up the cells for the implementation. And the appropriate survival of a certain amount of cells, can guard against the cells are being caused by pollution or other accidental changes in the cell dropping cells, played a role in cell care. In addition, you can also use the situation of cell cryopreservation to purchase, send, exchange and transport certain cells. When the cells are cryopreserved, the glycerol or dimethyl sulfoxide (DMSO), which is 5% ~ 15% of the final concentration, can reduce the freezing point of the solution, and in the case of slow freezing, the intracellular moisture , Eliminated the formation of ice crystals, thus preventing cell damage. The scale freezing rate starts at -1 to -2 ° C / min and can be accelerated when the temperature is below -25 ° C and can be directly injected into liquid nitrogen (-196 ° C) after -80 ° C.
  Commonly used screening agents are dimethyl sulfoxide (DMSO) and glycerol, they are non-toxic to cells, molecular weight, solubility, easy to penetrate cells.
  Implement pace
  1. Digest the cells and route the cell suspension to the centrifuge tube.
  2.Centrifuge at 10,000 rpm for 10 minutes, discard the supernatant.
  3. precipitation plus cover with the creation, counting, mediation to 5106 / ml or so.
  4. The suspension is divided into a cryopreservation tube, 1 ml per tube.
  5. Seal the frozen tube. If the ampoule is a flame seal, sealing must be strict, or wake up when prone to burst.
  6. Paste the label, specify the cell type, frozen date. Frozen tube tied to a metal weight and a string.
  7. Cool in the following order: room temperature → 4 ℃ (20 minutes) → refrigerator freezer (30 minutes) → low temperature refrigerator (-30 ℃ 1 hour) → gaseous nitrogen (30 minutes) → liquid nitrogen.
  Meticulous: manipulation should be alert to avoid liquid nitrogen frostbite. Liquid nitrogen regularly check, at any time to add, absolutely can not be volatile clean, as usual 30 liters of liquid nitrogen can use 1 to 1.5 months.
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