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Extended Hypothermic Storage of Human Hepatocytes Utilizing HypoThermosol

In the in vitro drug discovery/toxicity arena, primary hepatocytes serve as the gold standard cellular assay system due to their high sensitivity and diverse functional response.

In order to overcome the limited hepatocytes life span, we evaluated several commercially available hypothermic preservation media for their abilities to protect isolated human hepatocytes during hypothermic storage.

We investigated the temporal limits of these solutions in an attempt to extend storage while retaining a high degree of cell viability and function. Analysis of hepatocytes in the various hypothermic preservation media revealed that following 1-day of storage at 4°C, sample viability ranged from ~5% to ~60% (MF=5%, UW=35%, and HypoThermosol (HTS-FRS)= 60%). Extension of the preservation interval to 48 and 72 hours resulted in survival levels <15% in MF and UW whereas 40%(±4%) was achieved using HTS-FRS. Analysis of hepatocyte function demonstrated that along with the increase in viability in the HTS-FRS samples, there was high retention of hepatocellular function within the samples on a per cell basis.

 

These data indicate that hepatocyte storage can be extended 2-3 fold with HTS-FRS (HypoThermosol) while retaining viability and functionality levels at or above those attainable at 24 hours utilizing the current gold standard solution.

 


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