What do I need to prepare before cell cryopreservation?

Date:2022-06-01 10:17:33 Wednesday

Summary:In cell biology research, in order to better and long-term study cell biological function, it is necessary to preserve cells in good condition for future use. When the cells are directly frozen without any protective agent, the water in the cell and......

In cell biology research, in order to better and long-term study cell biological function, it is necessary to preserve cells in good condition for future use.

When the cells are directly frozen without any protective agent, the water in the cell and the external environment will form ice crystals, which can cause mechanical damage in the Cell spreader , increase in electrolytes, changes in osmotic pressure, dehydration, pH changes, protein denaturation, etc. Can cause cell death.

If a protective agent is added to the culture medium, the freezing point can be lowered. Under slow freezing conditions, intracellular water can be permeated out of cells before freezing. Storage at low temperatures below -130°C can reduce the formation of ice crystals.

When the cells are cryopreserved, they are out of the growth state and their cell characteristics are preserved, and they will re-enter the growth and division cycle when they are recovered. Before starting the experiment, put cryovials, 15 ml centrifuge tubes, pipettes, pipettes, pipette tips, etc. into a sterile ultra-clean workbench and irradiate them with UV light for 30 minutes. Use a ventilator for 3 minutes. Wipe the console and hands with 75% alcohol. Have an ice box ready.

Adjust the centrifuge to 800 rpm for 5 minutes. The water bath was adjusted to a constant temperature of 37 degrees. Take the cell complete medium, DMSO, trypsin, etc., and put it in a water bath to preheat.