Cryopreservation of human hepatocytes for clinical use.
Cryopreservation of hepatocytes is important for use both in research and for clinical application in hepatocyte transplantation. Cryopreservation causes damage to hepatocytes with the result that cell viability and function is reduced on thawing compared to fresh cells. There are many different protocols reported for freezing human hepatocytes mainly using DMSO as cryoprotectant. In this chapter the current detailed protocols used for cryopreservation and thawing of human hepatocytes for cell transplantation at the Cell Isolation Unit at King's College Hospital, London, are described. All procedures must be performed in a clean GMP environment using materials and reagents which are of pharmaceutical grade. The cryopreservation media is UW solution with added 300 mM glucose containing 10% DMSO and the thawing solution is EMEM containing 2% HSA. Freezing is performed in a controlled-rate freezer using a stepwise cooling programme.
Mitry
Ragai R
RR
Institute of Liver Studies, King's College Hospital and King's College London School of Medicine, London, UK.
Lehec
Sharon C
SC
Hughes
Robin D
RD
D000241:Adenosine; D000493:Allopurinol; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D005947:Glucose; D005978:Glutathione; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation; D006801:Humans; D007328:Insulin; D019188:Organ Preservation Solutions; D011887:Raffinose; D012709:Serum Albumin
D003451:Cryoprotective Agents; D007328:Insulin; D019188:Organ Preservation Solutions; D012709:Serum Albumin; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose; D004121:Dimethyl Sulfoxide