Cold storage of rat hepatocyte spheroids

Area of Science:

• Hepatology and regenerative medicine
• Cell preservation and biobanking

Background:

• Cell-based therapies for liver disease require viable hepatocytes for transplantation.
• Current cryopreservation methods lead to significant cell loss, hindering clinical application.
• Non-frozen cold storage offers a potential alternative for hepatocyte preservation.

Purpose of the Study:

• To optimize cold storage conditions for rat hepatocyte spheroids to maintain viability and function without freezing.
• To evaluate the efficacy of various storage solutions, including serum-free medium (SFM) with additives, for short-term hepatocyte preservation.

Main Methods:

• Rat hepatocytes were isolated and cultured into spheroids.
• Spheroids were cold stored at 4°C for 24 or 48 hours in different solutions (SFM, SFM + deferoxamine (Def), SFM + cyclosporin A (CsA), UW solution).
• Post-storage assessment included viability, gene expression, albumin production, and cytochrome P450/urea cycle enzyme activity.

Main Results:

• Cold storage induced injury, but deferoxamine (Def) significantly reduced this in both SFM and UW solutions.
• SFM + Def for 24 hours maintained high hepatocyte viability and function, approaching control levels.
• Storage in SFM alone or for 48 hours resulted in significant performance metric reductions.

Conclusions:

• A serum-free medium supplemented with deferoxamine enables hepatocyte spheroids to tolerate 24-hour cold storage with minimal viability and functionality loss.
• This optimized SFM + Def solution shows promise for improving cell-based liver therapies.
• Further research is needed to develop solutions for extended cold storage of hepatocyte spheroids.