Advances in Bioanalytics and Biomarkers

Combining 3D liver microtissues with lipid loading and lipidomics as a screening model for non-alcoholic fatty liver disease

Monday, February 5
4:30 PM - 5:00 PM
Location: 6E

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the world, affecting all racial, ethnic, and age groups without sex predilection. NAFLD is characterized by an excessive accumulation of lipids in hepatocytes (steatosis) and in combination with inflammatory processes (NASH) progressively develops into end stage liver disease, making it a major clinical concern.
Here, we describe a novel, screening-compatible human liver microtissue in vitro model for studying the etiology of steatosis and therapeutic strategies in a 3D configuration. The steatosis model is based on incubation with Oleate/Palmitate and displays a distinct and quantifiable accumulation of macro- and/or microvesicular lipid droplets within the hepatocytes. It maintains prolonged viability and liver-specific functionality in comparison to 2D cultures and can be produced in a 96-well SBS-compatible format.
Lipid accumulation was studied using fluorescent imaging followed by algorithmic analysis as well as by a novel LC/MS method for a full lipidomics analysis using unsupervised learning techniques.
Oleate as well as Palmitate induced a time- and concentration-dependent lipid accumulation, preferentially causing microvesicular (Oleate) or macrovesicular (Palmitate) steatosis. The highest lipid accumulation was observed after 7 days of Oleate treatment. The combination of both fatty acids in a physiological relevant 2:1 (Oleate:Palmitate) ratio resulted in a mixed phenotype. Lipidomics analysis confirmed increased concentrations of di- (18:1/18:1) and triglycerides (18:1/18:1/18:1) in microtissues upon treatment with Oleate or Oleate/Palmitate compared to medium and BSA control. In microtissues treated with Palmitate, increased concentrations of triglycerides (14:0/16:0/16:0) were observed. Lipidome principal component analysis allow for a clear distinction between the different treatment groups by corresponding clustering.

This 3D human liver microtissue model is particularly well-suited to study the formation as well as the prevention of steatosis by whole lipidome profiling, and is highly amenable for running comparisons to clinical samples. Moving beyond steatosis, the immune-competent status of these microtissues may even serve as starting point to study the etiology of NASH when combined with inflammatory stimuli.

Patrick Guye

Chief Scientific Officer
InSphero AG

The Chief Scientific Officer at InSphero AG, Dr. Guye possesses extensive experience in life sciences, engineering, business innovation & development, operations and new ventures consulting. He has held leadership positions in industry (Sanofi) and academia (Massachusetts Institute of Technology, Princeton University) with a focus on cell therapy, 3D cellular models/organoids, synthetic biology, human stem cells, biologics and small molecules development, and precision genomic engineering. Patrick earned his PhD in Molecular Infection Biology from the University of Basel.

Presentation(s):

Send Email for Patrick Guye


Assets

Combining 3D liver microtissues with lipid loading and lipidomics as a screening model for non-alcoholic fatty liver disease



Attendees who have favorited this

Please enter your access key

The asset you are trying to access is locked. Please enter your access key to unlock.

Send Email for Combining 3D liver microtissues with lipid loading and lipidomics as a screening model for non-alcoholic fatty liver disease