Pooling and re-freezing cells
Question: In order to get enough cells, we freeze individual aliquots, thaw them, pool the cells and then refreeze them. Will this harm the cells?
Answer: The short answer is 'yes'. All biospecimens (proteins, cells, tissues) suffer when they are repeatedly frozen and thawed. Try to avoid at all cost repeatedly freezing and thawing any biospecimen. For cell suspensions, freezing, thawig and then refreezing a cell will result in the majority of cells being dead. In fact repeatedly freezing and thawing cells is used to kill cancerous cells (when using cryosurgery to treat cancer). If, for whatever reason, you have to repeatedly freeze cells, it is best to have them culture for several days prior to refreezing. The culture period will give them a chance to recover fromt he nutrient deprivation, changes in membrane permeability and metabolic shifts experienced during freezing.
