Ozdener Mehmet Hakan, Rawson Nancy E
Monell Chemical Senses Center, Philadelphia, PA, USA.
Methods Mol Biol. 2013;945:95-107. doi: 10.1007/978-1-62703-125-7_7.
Establishment of primary and immortalized cultures of many cell types has facilitated efforts to understand the signals involved in proliferation and differentiation and yielded tools to rapidly assay new molecules targeting specific receptor pathways. Taste cells are specialized sensory epithelial cells which reside within taste buds on the lingual epithelium. Only recently have successful culturing protocols been developed which maintain essential molecular and functional characteristics. These protocols provide a tractable tool to examine the molecular, regenerative, and functional properties of these unique sensory cells within a controlled environment. The method involves an enzymatic isolation procedure and standardized culture conditions, and may be applied to either dissected rodent tissue or human fungiform papillae obtained by biopsy. Human fungiform cells can be maintained in culture for more than seven passages, without loss of viability and with retention of the molecular and biochemical properties of acutely isolated taste cells. Cultured primary human fungiform papillae cells also exhibit functional responses to taste stimuli indicating the presence of taste receptors and at least some relevant signaling pathways. While the loss of the three-dimensional structure of the intact taste bud must be taken into consideration in interpreting results obtained with these cells, this culture protocol provides a useful model for molecular studies of the proliferation, differentiation, and physiological function of mammalian taste receptor cells.
多种细胞类型的原代培养和永生化培养的建立,有助于人们了解增殖和分化过程中涉及的信号,并产生了用于快速检测靶向特定受体途径的新分子的工具。味觉细胞是位于舌上皮味蕾内的特殊感觉上皮细胞。直到最近才开发出成功的培养方案,能够维持其基本的分子和功能特性。这些方案提供了一种易于操作的工具,用于在可控环境中研究这些独特感觉细胞的分子、再生和功能特性。该方法包括酶分离程序和标准化培养条件,可应用于解剖的啮齿动物组织或通过活检获得的人菌状乳头。人菌状乳头细胞可以在培养中传代超过七次,而不会丧失活力,并保留急性分离的味觉细胞的分子和生化特性。培养的原代人菌状乳头细胞对味觉刺激也表现出功能反应,表明存在味觉受体和至少一些相关的信号通路。虽然在解释用这些细胞获得的结果时必须考虑完整味蕾三维结构的丧失,但这种培养方案为哺乳动物味觉受体细胞的增殖、分化和生理功能的分子研究提供了一个有用的模型。