Galactosylated cellulosic sponge for multi-well drug safety testing.

Area of Science:

• Hepatocyte biology and drug metabolism research
• Biomaterials and tissue engineering for drug discovery

Background:

• Hepatocyte spheroids maintain differentiated functions but aggregate and become hypoxic in extended cultures.
• Spheroid size limitations ( > 200 μm) hinder nutrient and drug access to inner cells.
• Current hepatocyte culture platforms face challenges in maintaining spheroid integrity and function for drug safety testing.

Purpose of the Study:

• To develop a novel scaffold for culturing functional 3D hepatocyte spheroids.
• To maintain spheroid size, prevent detachment, and support mature hepatocyte phenotype.
• To provide a scalable platform for drug safety testing applications.

Main Methods:

• Development of a thin, galactosylated cellulosic sponge scaffold for hepatocyte spheroid culture.
• Culture of hepatocytes within the macroporous scaffold in multi-well plates.
• Assessment of spheroid morphology, cell-cell interactions, polarity, and metabolic functions (e.g., cytochrome P450 activity).

Main Results:

• The galactosylated sponge supported rapid formation of hepatocyte spheroids with a mature phenotype.
• Tethered spheroids maintained 3D morphology, cell interactions, polarity, and metabolic/transporter functions.
• Cytochrome P450 activities were significantly elevated, and the sponge showed minimal drug absorption.

Conclusions:

• The galactosylated cellulosic sponge effectively supports and maintains functional hepatocyte spheroids.
• This platform offers a scalable alternative to current hepatocyte models for drug safety testing.
• The scaffold overcomes limitations of spheroid hypoxia and nutrient/drug access.