Cryopreservation of suckling XXXX hepatocytes.

@entity1707 @entity3175 @entity1133 @entity3733 @entity413
To determine the best and simplest method for cryopreservation of @entity1707 hepatocytes, we compared immediate cryopreservation with cryopreservation after short-term culture. Suckling @entity1707 hepatocytes were isolated by a modified 2-step in situ collagenase perfusion method, suspended in serum-free medium, and preserved for 10 da by two cryopreservation methods. Serial measurements were made of cell viability, LDH release, synthesis of protein, @entity1133 and @entity413 , @entity413 -6-phosphatase (G-6-Pase) activity, and @entity3175 transformation after thawing. These measurements were performed on both groups of cultured hepatocytes, and on freshly isolated hepatocytes, which served as a control. High viability (>95%)of thawed hepatocytes was obtained and maintained in both cryopreservation groups. There were no significant differences in cell viability, protein synthesis, @entity413 synthesis, G-6-Pase activity, or @entity3175 transformation between the two cryopreservation groups. In the immediate cryopreservation group, @entity1133 synthesis was less than in the group with cryopreservation after short-term culture. Protein synthesis, @entity413 synthesis, and @entity3175 transformation were lower in both cryopreserved groups than in the controls. The results showed that a protocol of immediate cryopreservation of hepatocytes in RPMI-1640 medium containing 10% @entity3733 , hormones, growth factors, and 10% newborn bovine serum, together with rate-controlled freezing and rapid thawing, provides indices of cell viability and function during subsequent serum-free culture that are comparable to hepatocytes cryopreserved after short-term culture, except for lower @entity1133 production. This simple procedure can be used in studies of bioartificial liver and hepatocyte transplantation.
