Cleavable cellulosic sponge for functional hepatic cell culture and retrieval

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Cell Biology

Background:

• Macroporous hydrogels support 3D cell culture but retrieving functional spheroids is challenging.
• There is a need for cleavable hydrogels that facilitate spheroid recovery for downstream applications.

Purpose of the Study:

• To develop and characterize a cleavable macroporous hydrogel sponge.
• To demonstrate its utility in supporting and enabling retrieval of functional hepatocyte spheroids.

Main Methods:

• Synthesis of a hydroxypropyl-cellulose-disulfide sponge conferring cleavability.
• Culture of HepaRG cells as spheroids within the macroporous hydrogel.
• Assessment of spheroid differentiation, function, purity, and retrieval using tris-(2-carboxyethyl)-phosphine (TCEP).

Main Results:

• The cleavable sponge supported hepatocyte spheroid formation and maintenance.
• HepaRG differentiation was accelerated (9 days vs. 28 days in 2D culture) with enhanced hepatocyte purity (92.9% albumin+ vs. 60%).
• Functional spheroids were rapidly retrieved within 30 minutes using TCEP, preserving cell viability and function.

Conclusions:

• A novel cleavable macroporous hydrogel sponge was successfully developed.
• This biomaterial facilitates efficient retrieval of viable and functional 3D hepatocyte spheroids.
• The technology holds promise for advancing 3D spheroid/organoid applications in research and drug development.