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Chemical Senses Advance Access originally published online on February 1, 2006
Chemical Senses 2006 31(3):279-290; doi:10.1093/chemse/bjj030
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© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Characterization and Long-Term Maintenance of Rat Taste Cells in Culture

Hakan Ozdener1, Karen K. Yee1, Jie Cao1, Joseph G. Brand1,2,3, John H. Teeter1 and Nancy E. Rawson1

1 Monell Chemical Senses Center, Philadelphia, PA 19104, USA, 2 Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA and 3 Veterans Affairs Medical Center, Philadelphia, PA 19104, USA

Correspondence to be sent to: Hakan Ozdener, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA. e-mail: hozdener{at}monell.org

Taste cells have a limited life span and are replaced from a basal cell population, although the specific factors involved in this process are not well known. Short- and long-term cultures of other sensory cells have facilitated efforts to understand the signals involved in proliferation, differentiation, and senescence, yet few studies have reported successful primary culture protocols for taste cells. Furthermore, no studies have demonstrated both proliferation and differentiation in vitro. In this study, we have developed an in vitro culture system to maintain and utilize rat primary taste cells for more than 2 months without losing key molecular and biochemical features. Gustducin, phospholipase C-ß2 (PLC-ß2), T1R3, and T2R5 mRNA were detected in the cultured cells by reverse transcriptase–polymerase chain reaction. Western blot analysis demonstrated gustducin and PLC-ß2 expression in the same samples, which was confirmed by immunocytochemistry. Labeling with bromo-2-deoxyuridine (BrdU) demonstrated proliferation, and a subset of BrdU-labeled cells were also immunoreactive for either gustducin or PLC-ß2, indicating differentiation of newly generated cells in vitro. Cultured cells also exhibited increases in intracellular calcium in response to several taste stimuli. These results indicate that taste cells from adult rats can be generated and maintained under the described conditions for at least 2 months. This system will enable further studies of the processes involved in proliferation, differentiation, and function of mammalian taste receptor cells in an in vitro preparation.

Key words: culture, gustducin, imaging, proliferation, taste receptor


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