Three-dimensional cultures of mouse submandibular and parotid glands: a comparative study

Abstract

Freshly isolated salivary cells can be plated on an extracellular matrix, such as growth factor-reduced Matrigel (GFR-MG), to induce the formation of three-dimensional (3D) structures. Cells grown on GFR-MG are able to form round structures with hollow lumina, capable of sustaining amylase expression. In contrast, cells grown on plastic do not exhibit these features. Our recent studies have used mouse parotid gland (PG) cells, grown on different extracellular matrices, as a model for acinar formation. However, PG cells were not able to respond to the secretory agonist carbachol beyond 5 days and did not sustain polarity over time, regardless of the substratum. An alternative option relies in the use of mouse submandibular glands (SMG), which are more anatomically accessible and yield a larger number of cells. We compared SMG and PG cell clusters (partially dissociated glands) for their ability to form hollow round structures, sustain amylase and maintain secretory function when grown on GFR-MG. The results were as follows: (a) SMG cell clusters formed more organized and larger structures than PG cell clusters; (b) both SMG and PG cell clusters maintained α-amylase expression over time; (c) SMG cell clusters maintained agonist-induced secretory responses over time; and (d) SMG cell clusters maintained secretory granules and cell–cell junctions. These results indicate that mouse SMG cell clusters are more amenable for the development of a bioengineered salivary gland than PG cell clusters, as they form more organized and functional structures.

Morphological Study on the Correlation of Prenatal and Postnatal Development between Mouse Parotid Salivary Gland and Tooth

“…8). These results are consistent with those from previous reports (Larsen et al, 2010;Leith et al, 2014;Melnick & Jaskoll, 2000;Redman, 1988;Riberiro et al, 2006), with slight differences in the timing of some of the details (Larsen et al, 2010;Leith et al, 2014;Ogawa et al, 2013;Okumura et al, 2012;Rego et al, 2016;Tucker, 2007). Of all the major salivary glands, the sublingual salivary gland is the first to form and development is already complete at birth (Chi, 1996;Poulsen et al, 1986;Redman, 1988).…”

“…This is different from the other major salivary glands. It is thought that complete formation of the parotid salivary gland is associated with the production and accumulation of secretory proteins in the acini at the final stage of postnatal development (Denny et al, 1997;Lawson, 1972;Leith et al, 2014;Redman & Sreebny, 1970). Important genetic changes can be observed during this period, which are consistent with the above results (Denny et al, 1997;Poulsen et al, 1986).…”

“…Of all the major salivary glands, the sublingual salivary gland is the first to form and development is already complete at birth (Chi, 1996;Poulsen et al, 1986;Redman, 1988). The submandibular and parotid salivary glands develop in a similar timeframe, although the submandibular salivary gland becomes fully developed slightly before the parotid gland (Chi, 1996;Larsen et al, 2010;Zhou et al, 2010) and forms a more organized structure than the parotid gland (Leith et al, 2014). However, there are conflicting reports about the details of the development of the salivary glands, and there are few reports on the mouse parotid salivary gland (Riberiro et al, 2006).…”

“…The acini, the secretory portion of the salivary gland, produce and secrete primitive saliva of which there are three types, serous, mucous and mixed saliva depending on the characteristics of the secretory cells (Amano et al, 2012;Chi, 1996). The parotid salivary gland is the largest of the salivary glands, and in the mouse the parotid glands consist of serous acinar cells that secrete an abundance of amylase (Amano et al, 2012;Leith et al, 2014;Tucker, 2007). Tooth development broadly comprises three stages, the bud stage, cap stage and bell stage and is a continuous process (Antonio, 2008).…”

Current trends in salivary gland tight junctions

“…Several recent studies have demonstrated that TJs can be formed from primary salivary in vitro, 110,[116][117][118][119] an advance which can be used to study barrier function of the salivary epithelium in a system that more closely resembles native tissue. Furthermore, the use of knockout mice for the various TJ proteins will also prove useful in determining the role of these proteins for in vivo salivary gland functioning.…”