Lumen formation in three-dimensional cultures of salivary acinar cells.

OBJECTIVE

Development of an artificial salivary gland will benefit patients with xerostomia after radiation therapy for upper respiratory cancer. The goal is to devise a three-dimensional (3D) culture system in which salivary cells differentiate into polarized acini that express essential biomarkers and directionally secrete alpha-amylase. Differentiated acini-like structures in a 3D biomaterial-based scaffold will mimic salivary gland functions.

STUDY DESIGN

Cells were seeded onto HA-based hydrogels containing PlnDIV peptide and allowed to differentiate into acini-like structures. Cell viability and phenotype were examined.

SETTING

Laboratory-based tissue procurement study.

SUBJECTS AND METHODS

Salivary gland tissue was obtained from patients undergoing surgery. Marker expression established the phenotype of salivary gland cells. Perlecan/HSPG2, an important component of the basement membrane, was highly expressed in salivary gland tissue. A culture system consisting of hyaluronic acid (HA) hydrogel and a coupled bioactive peptide derived from domain IV of perlecan (PlnDIV) was used. Prior studies demonstrated differentiation of acinar cells into lobular structures that mimicked intact glands when cultured on PlnDIV peptide-coated surfaces.

RESULTS

Lobular acini-like structures formed on hydrogels and expressed tight junction components such as zona occludens 1. Acini-like structures were stained for the presence of alpha-amylase. Live/dead staining revealed the presence of apoptotic cells in the center of the acini-like structures, indicative of lumen formation.

CONCLUSION

A novel system supporting acini-like assembly in a 3D culture system was established. Presence of biomarkers and secretion of salivary enzymes confirms functionality in vitro. Future experiments will test the 3D system in an animal model.