PhoenixBio is excited to be sponsoring a booth and attending the Microphysiological Systems World Summit 2025 (MPS 2025) in person in Brussels, Belgium from June 10th - 13th, 2025. Meet with our team at Booth #110 to learn more about PhoenixBio's capabilities and how we can help you improve lives through tomorrow's technologies.
The MPS Summit is the premier event to learn from the he latest scientific achievements, discuss the advances and challenges, and enable communication between young and newly interested scientists and pioneers of the MPS field.
While you're here, be sure to check our PhoenixBio poster presentation on June 12th in the morning and grab a chance to speak with our team at our booth!
Humanized Liver Related Abstracts
314 Positive effects of humoral factors secreted by cultured human hepatocytes from humanized liver chimeric mice on the maintenance of multiple hepatic functions in cryopreserved primary human hepatocytes
Cryopreserved primary human hepatocytes (cPHHs) are considered the gold standard in vitro tool for drug development. However, the limited supply and loss of multiple hepatic functions during culture remain unresolved challenges. Based on the isolation of human hepatocytes from cDNA-uPA/SCID mice based-humanized liver chimeric mice (PXB-mice), we have established a mass production system for human hepatocytes (PXB-cells) which have been providing them for over 10 years. We have also discovered that fibrin, plasma fibronectin, and other extracellular matrix (ECM) components derived from cultured PXB-cells provide a cell scaffold for cultured human hepatocytes through activation of the coagulation cascade. The ECM network promotes the re-establishment of cell polarity and supports the maintenance of hepatocyte functions. In this study, we investigated whether such an intrinsic factor-based self-maintenance system derived from cultured PXB-cells could be applied to cPHHs.
PXB-cells were isolated from PXB-mice and cultured with dHCGM (fresh medium; FM) as previously described. Culture supernatant was collected every two days from day 7 to 21 and stored as hepatocyte-derived conditioned medium (CM). cPHHs were cultured with FM, CM, and two commercial hepatocyte culture media (Lonza; Hepatocyte Culture Medium (HCM), Cellartis; Power Primary HEP medium (PM)) for 7 days. The impacts of each culture medium on mRNA expression, p450 activity, and cell morphology were examined with qPCR, P450-Glo (Promega), and immunostaining respectively. Efflux transporter activities were evaluated by the accumulation of Carboxydichlorofluorescein diacetate and Tauro-nor-THCA-24-DBD in the presence or absence of chemical inhibitors (Cyclosporin A, Bosentan, and Rifampicin).
CM-treated cPHHs exhibited higher p450 activities (CYP1A2, 2B6, 2C9, 3A4) compared to those cultured with other media. In comparison to cPHHs cultured with FM or other culture media, CM-treated cPHHs exhibited lower expression of TGFB and YAP signaling-related genes, as well as cholangiocyte markers, while demonstrating higher expression of hepatic marker genes. Immunostaining results indicated CM-treated cPHHs formed bile canaliculi, and the accumulation of fluorescent signals via efflux transporters was confirmed by co-incubation with chemical inhibitors.
These findings suggest that an intrinsic factors-based self-maintenance system could be applied for the preservation of multiple hepatocyte functions in cultured cPHHs.
Yuji Ishida1,2, Chihiro Yamasaki2, Go Sugahara1,2, Chise Tateno2 and Takeshi Saito1
1 Keck School of Medicine, University of Southern California, 2 Phoenixbio Co., Ltd.