Cryopreservation of Cynomolgus Monkey (Macaca fascicularis) Hepatocytes for Subsequent Culture and Protein Synthesis Studies

• Published in: In Vitro Cellular & Developmental Biology Vol. 26; no. 2; pp. 147 - 150
• Main Authors: Elena L. Sun, Danielle G. Aspar, Ulrich, Roger G., Melchior, George W.
• Format: Journal Article
• Language: English
• Published: Largo, MD Tissue Culture Association, Inc 01-02-1990; Society for In Vitro Biology
• Subjects: Albumins; Albumins - metabolism; Animal cells; Animals; Apolipoproteins A - metabolism; Biological and medical sciences; Biotechnology; Cells, Cultured; Centrifugation; Cryopreservation; Cryopreservation - methods; Cultured cells; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Hepatocytes; Liver; Liver - cytology; Liver - metabolism; Macaca fascicularis; Metabolism; Methods. Procedures. Technologies; Miscellaneous; Perfusion; Primates; Protein Biosynthesis; Viability; Cell culture; Albumin; Macaca fascicularis; Biosynthesis; Thawing; Apolipoproteins; Metabolism; Survival; Proteins; Vertebrata; Mammalia; Hepatocyte; Cryopreservation; Animal; Primates; Simioidea
• ISSN: 0883-8364; 2327-431X; 1475-2689
• DOI: 10.1007/bf02624105

A method is described for the preservation and subsequent recovery of hepatocytes obtained by collagenase perfusion of cynomolgus monkey (Macaca fascicularis) livers. The fresh cells are suspended in fetal bovine serum containing 10% dimethylsulfoxide and, using a microprocessor-controlled, liquid nitrogen freezing chamber and a specific cooling protocol, processed in such a way that they can be stored in liquid nitrogen for several months and still restored to active culture. When the cryopreserved cells were established in culture, they were found to actively synthesize and secrete both albumin and apolipoprotein A-I. That, taken together with morphologic evidence, was viewed as indication that the cells recovered in culture were in fact hepatocytes and not some other cell type from the monkey liver. The availability of this procedure for storing hepatocytes should contribute significantly to the efficient use of nonhuman primates as models with which to study hepatic metabolism.