The major challenge to cell cryopreservation is intracellular ice formation (IIF)

which can be reduced by controlling ice nucleation. In this review

the influence of ice nucleation on super-cooling and damage zone was analyzed. Physical

chemical

and biological methods have been applied to control ice nucleation during cryopreservation. The mechanisms of these methods were summarized and the influence of these methods on cell cryopreservation were discussed. Then it was proposed that biological and chemical methods should be superior methods of controlling ice nucleation. Additionally

the best ice seeding temperature was further proposed. At this temperature

ice crystals are initiated in the extracellular solution

which could result in minimal cell volume by reducing intracellularly active water properly

and then the incidence of IIF is reduced. The direction of ice seeding applied in the cell cryopreservation was pointed out.