Li Weijie, Yang Xi, Bi Wenyan, Song Liyong, Liu Baolin
Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology, Shanghai, China.
College of Chemistry and Chemical Engineering, Henan Polytechnic University, Jiaozuo, Henan, China.
Front Cell Dev Biol. 2025 Apr 28;13:1571198. doi: 10.3389/fcell.2025.1571198. eCollection 2025.
Developing bioartificial liver and hepatocyte transplantation technology causes increasing hepatocyte cell demand. Effective long-term hepatocyte cell preservation methods are necessary to promote. Progressive cooling is a key preservation technology for cell banks. However, the cell solution needs to be supercooled in a slow freezing process. The high degree of supercooling possibly induces uncontrollable intracellular ice formation. This work designs an ultrasonic ice-seeding system for L-02 hepatocyte cell preservation, reducing supercooling and improving cell survival rate. The effect of ultrasonic intensities on the hepatocyte's survival rate was investigated and optimized. The results prove the calorimetric method can efficiently measure the ultrasonic intensity dissipated in the hepatocyte cell preservation solution. When the ultrasonic intensity is 0.0329 W/cm ∼ 0.4316 W/cm, the hepatocyte survival rate is over 90%. There is no significant difference between experiment groups (p < 0.05) when the ultrasonic intensity is larger than 0.4316 W/cm. The hepatocyte cell survival rate reduced significantly with the increase of ultrasonic intensity. The 7-day hepatic function indicator experiment results indicate that the ultrasonic ice seeding has the weakest impact on hepatocyte cells in the four groups. The secretion of urea, albumin and glucose proved that ultrasonic ice seeding technology does not affect cell secretion and has an enormous advantage in cryopreservation. It can be widely applied to cell freezing fields.
生物人工肝和肝细胞移植技术的发展导致肝细胞需求不断增加。因此，需要有效的长期肝细胞保存方法来推动这一发展。程序降温是细胞库的关键保存技术。然而，在慢速冷冻过程中，细胞溶液需要过冷。高度过冷可能会导致不可控的细胞内冰晶形成。本研究设计了一种用于L-02肝细胞保存的超声引晶系统，以减少过冷现象并提高细胞存活率。研究并优化了超声强度对肝细胞存活率的影响。结果表明，量热法能够有效地测量肝细胞保存溶液中耗散的超声强度。当超声强度为0.0329 W/cm至0.4316 W/cm时，肝细胞存活率超过90%。当超声强度大于0.4316 W/cm时，各实验组之间无显著差异（p < 0.05）。随着超声强度的增加，肝细胞存活率显著降低。为期7天的肝功能指标实验结果表明，超声引晶对四组中的肝细胞影响最弱。尿素、白蛋白和葡萄糖的分泌证明，超声引晶技术不影响细胞分泌，在冷冻保存方面具有巨大优势。它可广泛应用于细胞冷冻领域。
