Freezing Cells

If you want columns a and b to remain still as you scroll sideways through your data click column c to select it.

Freezing cells. It s also called cryosurgery or cryoablation. Thawing cells 1 remove cells from the tank and thaw. During cryotherapy treatment the doctor freezes the cancer cells to kill them. Place vials on wet ice or in a 4 c refrigerator and start the freezing procedure within 5 minutes.

Follow this protocol to. Cryotherapy uses extreme cold to destroy cancer cells. The shape of cells gets distorted by ice crystals causing mechanical damage. When you have started a new cell line it is a good policy to freeze down a good portion of the cells for use at a later date.

Select view freeze panes freeze panes. If you want to keep rows 1 2 and 3 in place as you scroll down through your data click row 4 to select it. Alternatively place the cryovials containing the cells in an isopropanol chamber and store them at 80 c overnight. For the greatest cell viability it is important to freeze the cells slowly.

Cells are frozen slowly at 1 c min. For example if you thaw a vial of cos cells to carry the cell line you will eventually split the cells into 10 15 plates. Cryotherapy is called a local treatment which means that it only treats the area where you have treatment. Transfer frozen cells to liquid nitrogen and store them in the gas phase above the liquid nitrogen.

3 check your cells. Frozen cells must connect to the top or left edge of the spreadsheet. The solutes in the unfrozen water get concentrated causing chemical and osmotic damage. Immediately transfer the cells to a large volume of pre warmed cell growth media.

Remember to spray everything down with 70 ethanol and wipe it with kimwipes before moving into the bsc. Aliquot into cryogenic storage vials. About 24 hours after. As ice crystals form cells get packed into small pockets of unfrozen liquid.

Methods for freezing cells general warm cell medium and reagentpack reagents to 37 c in the water bead bath. Once you have done this freeze down a large portion of the plates in cryovials. Freeze rows or columns select the cell below the rows and to the right of the columns you want to keep visible when you scroll. Freeze the cells in a controlled rate freezing apparatus decreasing the temperature approximately 1 c per minute.

Thaw cell freezing medium and keep it at 2 8 c until use. Resuspend cells in freezing medium to a concentration of 1 x 10 7 to 5 x 10 7 cells ml for serum containing medium or 0 5 x 10 7 to 1 x 10 7 cells ml for serum free medium.