Abstract
Induced pluripotent stem cell derived hepatocytes (IPSC-Heps) have the potential to reduce the demand for a dwindling number of primary cells used in applications ranging from therapeutic cell infusions to in vitro toxicology studies. However, current differentiation protocols and culture methods produce cells with reduced functionality and fetal-like properties compared to adult hepatocytes. We report a culture method for the maturation of IPSC-Heps using 3-Dimensional (3D) collagen matrices compatible with high throughput screening. This culture method significantly increases functional maturation of IPSC-Heps towards an adult phenotype when compared to conventional 2D systems. Additionally, this approach spontaneously results in the presence of polarized structures necessary for drug metabolism and improves functional longevity to over 75 days. Overall, this research reveals a method to shift the phenotype of existing IPSC-Heps towards primary adult hepatocytes allowing such cells to be a more relevant replacement for the current primary standard.
Highlights
With uses in drug screening, toxicology studies, cell-based therapies, and in vitro disease modeling, primary human hepatocytes (PHHs) are in high demand
We began by investigating the importance of cell-cell junction maintenance during the transfer of the cells from 2D to 3D culture (Figure 1)
Media samples were taken at day 25, 35, and 45 and were subjected to immunoassays in order to quantify the secretion of human serum albumin, alpha-1-antitrypsin (A1AT), and alphafetoprotein (AFP), which marks fetal hepatocytes
Summary
With uses in drug screening, toxicology studies, cell-based therapies, and in vitro disease modeling, primary human hepatocytes (PHHs) are in high demand. Several studies have demonstrated that culturing isolated PHHs in a 3D format averts many effects of dedifferentiation and can partially reverse this process in cells that have been cultured for short term in monolayer (2-Dimensional conditions) [7] Such 3D cultures have been shown to return the function of several xenobiotic metabolizing enzymes to in vivo levels [8,9], reestablish cellular polarization and canalicular structure [9,10], and maintain other liver specific functions such as albumin secretion, glycogen synthesis, and lipid storage [7,9]. 3D culture systems currently available are often unwieldy and overly complex, leading to poor reproducibility and restricting use to a few labs with highly specialized equipment Such methods, often based upon embryoid body differentiation, are not compatible with high throughput screening and remain difficult to apply to IPSC-Heps, which require long term, reproducible culture for functional differentiation and subsequent application in research and industry
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
