Neural influence on salispheres development

Objectives: Salispheres are spheroid hetero-cellular non-adherent clusters, which form in vitro from dissociated salivary gland cells. Selected cells isolated from salispheres contribute decisively to the recovery of experimental-induced atrophic salivary glands. However, the properties of salispheres change after five days in culture, seriously diminishing their inductive potential. It is critical to understand why salispheres become dormant in tissue culture. The main aim of this study is to assess whether neural factors affect salispheres formation and development.
Methods: We harvested submandibular glands from adult female mice (ICR strain) under terminal anesthesia and processed them by mechanical and enzymatically digestion. The morphology of cells has been examined for ten days from plating under phase contrast microscope (cell number, size, morphology and aggregating properties). Neural mediators (atropine, carbachol, substance P and VIP) have been selectively added to the cell culture.
Results: Salispheres development was influenced by different neural mediators. Most salispheres formed in an atropine-enriched microenvironment were slightly smaller compared to the control (57.03mm vs. 76.41 average diameter in Day 4). Adherent properties of salispheres were increased in the substance P-treated group (approx. 10% of Day 4 salispheres had already stuck), and occasional mixed aggregates of salispheres and adherent cells were encountered.
Conclusions: This study showed that neural mediators influence the developing pattern of salispheres formation in vitro. By modulating the culture microenvironment, the results of using salispheres in salivary gland regeneration might be enhanced.