Category: Cellular Technologies

1250-A - A novel maintenance medium extends the lifespan and enables long-term applications for both human primary hepatocytes and human pluripotent stem cell-derived hepatocytes in conventional 2D cultures

Wednesday, February 7, 2018
11:30 AM - 12:30 PM

Human primary hepatocytes are considered the gold standard for in vitro model systems of liver function for drug development, toxicity assessment, and metabolic disease research; however, their rapid loss of cell viability in conventional 2D culture limits their utility in these applications. Human induced pluripotent stem (hiPS) cell-derived hepatocytes have potential as a better in vitro model if they possess a relevant usage window and functionality—but this is challenging to accomplish.


Addressing these problems, our newly developed hepatocyte maintenance medium enables the culture of cryopreserved human primary hepatocytes or hiPS cell-derived hepatocytes for four or two weeks, respectively, with maintained viability and stable activities of several key cytochrome P450 enzymes (CYPs). Multiple analyses on cryopreserved hiPS cell-derived hepatocytes, including RT-qPCR, immunostainings, functional assays such as albumin secretion, and CYP activity assays demonstrate mature features and high functionality. Importantly, the hiPS cell-derived hepatocytes show expression of the essential genes of the drug-metabolizing machinery, such as CYPs, phase II enzymes, and transporters. 


An extended in vitro culture time for hepatocytes enables chronic toxicity testing. We show that hiPS cell-derived hepatocytes can be exposed to known hepatotoxins for up to 14 days. Cells respond as expected to these toxic compounds, demonstrating their utility for chronic toxicity studies. The hiPS cell-derived hepatocytes also respond to insulin, and they can take up and store low-density lipoproteins and fatty acids.


The novel maintenance medium presented here maintains the viability and functionality of cryopreserved human primary hepatocytes and hiPS cell-derived hepatocytes from multiple lines for a much longer time than existing commercially available hepatocyte maintenance media. We hope that the increased assay window of functional hepatocytes in 2D cultures will empower new areas of liver research and applications.

Liz Quinn

Associate Director for Stem Cell Marketing
Takara Bio USA, Inc.
Mountain View, CA

Dr. Liz Quinn is the Associate Director for Stem Cell Marketing at Takara Bio USA following nearly 20 years of research and commercial experience in the life science and biotechnology industries. Dr. Quinn joined Clontech Laboratories, Inc. in 2001 and held varied responsibility within Operations, R&D and Marketing. In 2008, she joined DiscoverX Corporation and managed the GPCR drug discovery business. After leaving DiscoverX in 2014, she served as founder and CEO for a new life science and drug discovery-focused marketing consulting company based in Northern CA. Then in 2015, Liz joined Takara Bio USA where she has focused on launching a new stem cell product portfolio which includes human iPS and ES-derived lines and culture systems for gene editing. In addition to her current role, Liz is a board member of ELRIG, the European Laboratory Research and Innovation Group, and has served as their Scientific Programme Director since 2014.