Characterization of Murine Autologous Salivary Gland Graft Cells: A Model for Use with an Artificial Salivary Gland

Abstract

The purpose of this study was to examine the growth and key functional abilities of primary cultures of salivary epithelial cells toward developing an artificial salivary gland. Cultures of epithelial cells originating from submandibular glands of BALB/c mice were established. Parenchymal cells were isolated by a Percoll gradient technique and thereafter seeded on irradiated NIH 3T3 fibroblasts serving as a feeder layer. The isolated cells were termed autologous salivary gland epithelial (ASGE) cells and could be cultivated for at least five passages (time limit of experiments). ASGE cells presented the typical organizational behavior of epithelial cells and electron microscopy, as well as immunostaining for cytokeratins, confirmed their epithelial origin. Furthermore, measurements of transepithelial resistance and water permeability indicated the ability of the ASGE cells to form a functional epithelial barrier. This study suggests that primary salivary epithelial cells can be obtained that exhibit critical characteristics needed for use with an artificial secretory device.

Primary Sjögren’s Syndrome: Current Pathophysiological, Diagnostic and Therapeutic Advances

“…Tissue engineering of salivary glands utilizes cells, biodegradable scaffold, and signals to regenerate tissues. Since the pioneer work reporting the culture of salivary epithelial cell culture (Brown 1974), several culture procedures have been described (Horie et al, 1996;Aframian et al, 2004;Joraku et al, 2005;Tran et al, 2006). A multipotent stem cell population has been discovered in human adult salivary glands (Okumura et al, 2003;Hisatomi et al, 2004;Kishi et al, 2006), but their potential for engineering salivary glands has not been proven.…”

Isolation and Cultivation of Integrin α6β1–Expressing Salivary Gland Graft Cells: A Model for Use with an Artificial Salivary Gland

“…The isolated cells were inoculated into 25-cm 2 Falcon flasks pre-seeded with 2Â10 5 lethally irradiated (50 Gy) NIH 3T3 mouse fibroblasts serving as a feeder layer, as previously described, 6 or into 10 mg/cm 2 of rat collagen type I pre-coated flasks (Sigma). Parenchymal cells were maintained in a 1:1 mixture of Dulbecco's modified Eagle medium and Ham's F-12 supplemented with 2 mM glutamine, penicillin (100 units/mL), streptomycin (100 units/mL), and amphotericin (0.25 mg/mL) (Biological Industries).…”

“…2,3 In recent years, we and others have concentrated on methods for regenerating the SGs' normal function in these patients. [4][5][6] Such regeneration is based on the assumption that autologous SG graft cells may be isolated before initializing irradiation therapy, cultivated, and preserved during the irradiation period. Upon therapy completion, cells would then be implanted into the irradiated gland, replacing the damaged functional cells.…”

“…In a scenario in which SG residual tissue cannot be used as a scaffold for the implantation, cells would be used in the context of an artificial SG device in anticipation of resuming secretory ability. [4][5][6] Previous reports have described several methods to establish primary cultures of SG epithelial cells. [7][8][9][10][11][12][13] However, cell division capability was restricted to several passages, and differentiation was almost impossible.…”

Prospect of functional salivary gland enhancement with biomedical strategies for head and neck cancers

“…Since then, several culture procedures have been published, initially by use of feeder cells [29] , and more recently using a serum-free medium for epithelial cells [30,31] .…”