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J Tissue Eng Regen Med. 2019 May 31. doi: 10.1002/term.2908. [Epub ahead of print]

Cell culture of differentiated human salivary epithelial cells in a serum-free and scalable suspension system: the Salivary Functional Units model.

Author information

1
McGill Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, Montreal, QC, Canada.
2
Department of Biomedical Sciences, University of Cagliari, Cittadella Universitaria di Monserrato, Cagliari, Italy.
3
Department of Anatomy and Histology, Faculty of Medicine, University of Jordan, Amman, Jordan.
4
Department of periodontics, School of dentistry, University of Montreal, Montreal, QC, Canada.
5
Department of Otorhinolaryngology-Head & Neck Surgery, Pusan National University School of Medicine, Pusan, South Korea.
6
Department of Anatomy and Cell Biology, McGill University, Montreal, QC, Canada.
7
Department of Otolaryngology- Head &Neck Surgery, McGill University, Montreal, QC, Canada.
8
Department of Oral and Maxillofacial Surgery, McGill University, Montreal, QC, Canada.

Abstract

Saliva aids in digestion, lubrication, and protection of the oral cavity against dental caries and oropharyngeal infections. Reduced salivary secretion, below an adequate level to sustain normal oral functions, is unfortunately experienced by head and neck cancer patients treated with radiotherapy, and by patients with Sjögren's syndrome. No disease-modifying therapies exist to date to address salivary gland hypofunction (xerostomia, dry mouth) because pharmacotherapies are limited by the need for residual secretory acinar cells, which are lost at the time of diagnosis, while novel platforms such as cell therapies are yet immature for clinical applications. Autologous salivary gland primary cells have clinical utility as personalized cell therapies, if they could be cultured to a therapeutically useful mass while maintaining their in vivo phenotype. Here, we devised a serum-free scalable suspension culture system that grows partially-digested human salivary tissue filtrates comprising of acinar and ductal cells attached to their native extracellular matrix components while retaining their 3D in vivo spatial organization; we have coined these salivary spheroids as "Salivary Functional Units (SFU)". The proposed SFU culture system was sub-optimal, but we have found that the cells could still survive and grow into larger salivary spheroids through cell proliferation and aggregation for 5 to 10 days within the oxygen diffusion rates in vitro. In summary, by using a less disruptive cell isolation procedure as the starting point for primary cell culture of human salivary epithelial cells, we demonstrated that aggregates of cells remained proliferative and continued to express acinar and ductal cell-specific markers.

KEYWORDS:

Dry mouth syndrome; Human primary epithelial cells; Salivary Functional Units; Salivary acinar cells; Salivary gland regeneration; Serum-free culture; Suspension culture; Xerostomia

PMID:
31151134
DOI:
10.1002/term.2908

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