The Cell Cultivation Study Of Human And Mouse Taste Buds

Background: Taste is a feeling caused by external stimulations act on the intraoral taste buds, and it’s one of the most important ways for people and other animals to receive external stimulations. Taste plays an important role in the maintenance of energy balance and homeostasis in the body. In clinical, many diseases (such as tongue cancer) and treatments (such as head and neck cancer radiotherapy) can result in dysgeusia, and even lead to losing the sense of taste. Compared with the research of visual、audition and olfactory, the knowledge of taste is very limited. So we can not take effective measures to timely intervene and treat the dysgeusia in clinical. It will greatly accelerate the development of the study of taste, if we can study taste bud cells which cultivated in vitro. Yet few studies have reported successful primary cultivating protocols for taste cells. Therefore, to establish a better and steadier method to cultivate human and ICR mouse taste bud cells is very necessary.Objective:To establish a better and steadier method to cultivate human and ICR mouse taste bud cells.Method:1. Determine the location of the taste buds on the back of the tongue by using HE staining and Immunohistochemistry methods.2. Collecte normal tongue mucosa from extended resection surgery of tongue cancer and ICR mouse, which contains taste papilla. Use tissue culture and acquisition of viable taste cells by enzymatic treatment to cultivate taste bud cells.3. Use the specific proteins of Taste buds to identify cultivated cells.Result1. By HE staining, it is found that taste buds are contained on the tongue fungiform papillae、circumvallate papillae and foliate papillae, while is not contained on the filiform papillae.2. Immunohistochemistry and immunofluorescence was observed for G‐protein alpha‐gustducin in taste bud and in about20%‐30%of taste bud cells. Immunohistochemistry and immunofluorescence for CK8was also observed in taste bud and about all of the taste bud cells.3. Use tissue cultivation and acquisition of viable taste cells by enzymatic treatment to taste bud cells cultivate. These results indicate that taste cells from human and ICR mouse can be maintained under the described conditions for at least15‐18days.4. In this study，the taste bud cells denervation of gustatory nerves，we maintained in cultivation for more than18days without loss of viability and while retaining express some taste cell–specific proteins.Conclusion:1. We established a method for enzymatically isolating human cells and ICR mouse from fungiform papillae and maintaining them in cultivation for short term without loss of viability and retaining express some taste cell–specific proteins.2. We established a method for taste bud cells from circumvallate papillae tissue cultivation and maintaining them in cultivation.