3D Spheroid Cultures for Salivary Gland Tissue Engineering: Effects of Fibroblast on Epithelial Cell Function

TL;DR

This study explores how combining epithelial cells and fibroblasts in 3D spheroids can help create salivary gland-like tissues, with a specific cell ratio showing the best results.

Contribution

The study identifies an optimal 2:1 epithelial to fibroblast ratio for maintaining salivary gland epithelial cell function in 3D spheroids.

Findings

A 2:1 ratio of epithelial cells to fibroblasts preserved epithelial cell phenotypes and structural integrity.

The optimal ratio reduced apoptosis and senescence markers while increasing progenitor marker expression.

These spheroids may serve as a model for salivary gland tissue engineering.

Abstract

Three-dimensional (3D) spheroid cultures are crucial for modeling salivary gland (SG) morphogenesis and advancing regenerative medicine. This study evaluated the effects of varying ratios of mouse SG-derived epithelial cells co-cultured with human dermal fibroblasts (hDFs), identifying a 2:1 ratio (spheroids containing 67% EpCAMpos cells with 33% hDFs) as optimal for preserving native SG-derived epithelial cell phenotypes. At this ratio, 67% EpCAMpos spheroids maintained structural integrity and demonstrated a significant reduction in apoptosis and senescence markers, specifically, cleaved caspase-3 (Cc3) and Serpine1, alongside an enhanced expression of the progenitor marker Keratin 5 (KRT5). This highlights the pivotal role of fibroblasts in supporting epithelial cell function in 3D cultures. These spheroids provide a useful model for developing SG tissues that closely mimic physiological properties. Despite promising results, these findings are preliminary and require further validation under diverse conditions and across different SG models.

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