# 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.

## Key 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.