Authors

D Whaley¹; K Daymar¹; C Luong¹; I Xu¹; G Peixoto¹; J Mönch²; M Alexander¹; RP Witek³; J Lakey¹; T Bates⁴; 
¹ University of California Irvine, United States;  ² Nordmark Pharma GmbH, Germany;  ³ Ambys Medicines, United States;  ⁴ AMS Biotechnology (Europe), UK

Discussion

Hepatocyte transplantations for acute liver failure and metabolic liver disorders represent a potential alternative to traditional orthotopic liver transplantation (OLT). Additionally, cultured primary hepatocytes are an important tool for in vitro pharmacological studies. Advances in clinical hepatocyte transplantation and drug studies depend on the development of standardized isolation and culturing protocols as well as high-quality enzymes for liver tissue digestion. The isolation of primary hepatocytes for clinical applications must comply with current Good Manufacturing Practices (cGMP) regulations which may include the use of digestion enzymes produced to cGMP standards.

In this study, we compare VitaCyte^® GMP-grade Collagenase MA in combination with BP Protease to Nordmark^® Collagenase NB 6 GMP-Grade. Additionally, we compare non-GMP-grade Nordmark^® Collagenase NB 4G Proved Grade to GMP-grade Nordmark^® Collagenase NB 6 to identify a high-quality cost-effective alternative for hepatocyte isolation in non-clinical research applications. Our results indicated that Nordmark^® NB 6 and VitaCyte^® MA + BP blend produced hepatocytes with comparable pre-purification viable yields and viability. Similarly, non-GMP-grade Nordmark^® NB 4G furnished primary hepatocytes with pre-purification yields and viabilities not significantly different from GMP-grade Nordmark^® NB 6. This suggests that GMP-grade Nordmark^® NB 6 is a viable alternative to GMP-grade VitaCyte^® MA+BP enzyme blend for primary hepatocyte isolation, and that non-GMP-grade Nordmark^® NB 4G is an affordable alternative for hepatocyte application in research. GMP-grade VitaCyte^® and Nordmark^® are manufactured to cGMP standards and can be utilized in the development of protocols intended for clinical applications.