Primary culture of mammalian taste epithelium.
Primary culture of mammalian taste epithelium.
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哺乳动物味觉上皮的原代培养。
DOI：
10.1007/978-1-62703-125-7_7
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Rawson,NancyE
中科院分区：
文献类型：
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作者：
Ozdener,MehmetHakan;Rawson,NancyE
许多细胞类型的原代和永生化培养物的建立促进了理解增殖和分化所涉及的信号的努力，并产生了快速检测针对特定受体途径的新分子的工具。味觉细胞是专门的感觉上皮细胞，位于舌上皮的味蕾内。直到最近才开发出成功的培养方案，以保持基本的分子和功能特征。这些协议提供了一种易于处理的工具，可以在受控环境中检查这些独特感觉细胞的分子、再生和功能特性。该方法涉及酶分离程序和标准化培养条件，并且可应用于解剖的啮齿动物组织或通过活检获得的人类真菌状乳头。人蕈状细胞可以在培养物中维持七代以上，而不会丧失活力，并且保留了敏锐分离的味觉细胞的分子和生化特性。培养的原代人蕈状乳头细胞还表现出对味觉刺激的功能反应，表明味觉受体和至少一些相关信号传导途径的存在。虽然在解释这些细胞获得的结果时必须考虑完整味蕾三维结构的损失，但该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有用的模型。
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.