Cryopreserved human hepatocytes in suspension are a convenient high throughput tool for the prediction of metabolic clearance

• Published in: European journal of pharmaceutics and biopharmaceutics Vol. 63; no. 3; pp. 347 - 355
• Main Authors: Jouin, Delphine, Blanchard, Nadège, Alexandre, Eliane, Delobel, Frédéric, David-Pierson, Pascale, Lavé, Thierry, Jaeck, Daniel, Richert, Lysiane, Coassolo, Philippe
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.07.2006; Elsevier Science
• Subjects: Aged; Aged, 80 and over; Animals; Biological and medical sciences; Clearance prediction; Cryopreservation; Cryopreserved; Female; General pharmacology; Hepatocytes - metabolism; High throughput; Human hepatocytes; Humans; Male; Medical sciences; Metabolic Clearance Rate; Middle Aged; Miniaturisation; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Rats; Rats, Wistar; Suspension; Suspensions; Miniaturisation; High throughput; Clearance prediction; Suspension; Human hepatocytes; Cryopreserved; Human; Pharmaceutical technology; Prediction; Clearance; Metabolism; In vitro; Hepatocyte; Dosage form; Pharmacokinetics
• ISSN: 0939-6411; 1873-3441
• DOI: 10.1016/j.ejpb.2006.01.014

Hepatocyte assays, routinely used to assess the metabolic stability of new chemical entities, were recently improved by using hepatocytes in suspension instead of primary cultures [N. Blanchard, L. Richert, B. Notter, F. Delobel, P. David, P. Coassolo, T. Lavé, Impact of serum on clearance predictions obtained from suspensions and primary cultures of rat hepatocytes, Eur. J. Pharm. Sci. 23 (2004) 189–199]. The aim of the present study was to investigate miniaturising the suspension assay by using cryopreserved human hepatocytes, i.e., 150,000 cells/well in 96-well plates, to predict hepatic clearance (CL H) in order to increase compound throughput and decrease cost and tissue requirements. For this, an evaluation was first carried out with rat hepatocytes. Then, human hepatocytes from various donors were used under these predetermined conditions, either immediately after isolation, either after a 20-h-cold storage period in UW or after cryopreservation. The values of CL int and CL H determined using human hepatocytes in suspension in 96-well plates, immediately after isolation, after cold storage or after cryopreservation, were comparable to those obtained with hepatocytes in primary culture. In particular, the use of cryopreserved human hepatocytes in suspension in a 96-well format appeared to be largely satisfactory as a tool for screening and ranking of compounds in the early phase of the drug discovery process.