Characterization of Gastrospheres Using 3D Coculture System

Area of Science:

• Cell Biology
• Gastroenterology
• Biomedical Engineering

Background:

• Understanding the molecular mechanisms of gastric disorders and regeneration requires an in vitro model that accurately mimics the in vivo gastric microenvironment.
• Existing models may not fully capture the complex cellular interactions and structural organization of the gastric tissue.

Purpose of the Study:

• To develop and characterize a novel three-dimensional (3D) coculture system, termed gastrospheres, for studying gastric disorders and regeneration.
• To assess the ability of gastrospheres to recapitulate the in vivo gastric microenvironment.

Main Methods:

• Primary human gastric epithelial (HGE3) and stromal (HGS12) cells were isolated from endoscopic biopsies and expanded.
• Cells were cultured in a 3D system (gastrospheres) using agarose-coated plates and maintained in a rotary cell culture system (RCCS-4) for up to 28 days.
• Gastrospheres were characterized using immunocytochemistry (cytokeratins, vimentin, PCNA, nestin) and electron microscopy (SEM, TEM).

Main Results:

• Gastrospheres exhibited a distinct structure with cytokeratin-positive epithelial cells on the outer layer and vimentin-positive stromal cells in the center.
• Proliferating cell nuclear antigen (PCNA)-positive cells were observed in peripheral and intermediary regions, indicating active cell division.
• Electron microscopy revealed cohesive gastric cells, differentiated mucous/zymogenic-like cells, and central stromal structures, alongside extracellular matrix deposition.

Conclusions:

• The developed 3D gastrosphere model effectively recapitulates key features of the in vivo gastric microenvironment.
• Gastrospheres provide a valuable platform for investigating gastric pathophysiology, cellular interactions, and potential therapeutic strategies for gastric disorders and regeneration.