During the slow freezing process

the cell solution is super-cooled

which may cause uncontrollable intracellular ice formation. Ultrasonic ice seeding can reduce super-cooling and improve the cell survival rate. An ultrasonic ice seeding device was designed for the ice seeding preservation of L-02 hepatocytes

and the influence of different ultrasonic intensities

different pre-cooling temperatures

and different volume concentrations of Me2SO on the survival rate of hepatocytes were explored. The results show that when Me2SO with a volume concentration of 0%–10% is used as a cryoprotectant

the use of ultrasonic ice seeding operation can significantly improve the survival rate of hepatocytes during freezing. When the ultrasound intensities are 0.032 9 W/cm2~0.431 6 W/cm2

the hepatocyte survival rate is above 90%

and there is no significant difference between the groups (p<0.05). When the ultrasound intensity is greater than 0.431 6 W/cm2

the hepatocyte survival rate decreased significantly; ultrasonic ice seeding with Me2SO at a volume concentration of 5% at ﹣7 ℃ and an ultrasonic intensity of 0.1041 W/cm2 results in the highest survival rate of hepatocytes

which is (97±2.6)% and is not significantly different from the fresh group. Therefore

ultrasonic ice seeding can significantly improve the survival rate of hepatocytes and reduce the amount of Me2SO in the solution.