Name: isolation and culture of human fungiform taste papillae cells
Uploaded: 2012-05-17T12:00:00
Description: Watch this Scientific Journal Video about Isolation and Culture of Human Fungiform Taste Papillae Cells at JoVE.com

我们感谢艾米迈尔斯和ESI Quayson技能和帮助。这项工作是支持部分由NSF 0216310和奇华顿公司授予（净入学率）。

1。获得人力蕈状味觉​​乳头<ol><li>取出四到八个舌头的前部分采用弧形弹簧microscissors背表面的菌状味觉乳头。 </li><li>立即进入隔离解决方案的地方（ 碳酸氢钠 26毫米，2.5毫米的NaH 2 PO 4计 ，葡萄糖20毫米，65毫米氯化钠，氯化钾20毫米，1毫米EDTA在无核酸酶水溶解和过滤消毒）。 </li></ol> 2。人菌状的味道乳头消解人蕈状味觉细胞乳...

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我们一直保持着味道从人类的蕈状乳头细胞获得超过8通道，横跨12个月的期限。这些文化产生新的细胞，其中许多在体外成熟的阶段，他们表示，除了味觉受体细胞的关键分子和生理属性的几个指标，成熟的味蕾细胞类型。存在gustducin和PLC-β2在培养的细胞亚群的免疫反应是与这一结论相一致。最重要的是，某些细胞对甜和苦味刺激的应用与细胞内钙离子的短暂增加，这是一个功能分离的味...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td></tr><tr><td> MCDB上123 </td><td> Sigma-Aldrich公司</td></tr><tr><td> iscove的DMEM培养基</td><td>敏达</td></tr><tr><td>弹性蛋白酶</td><td> Sigma-Aldrich公司</td></tr><tr><td>蛋白酶E </td><td> Sigma-Aldrich公司</td></tr><tr><td>弹性蛋白酶</td><td>沃辛顿</td></tr><tr><td>大豆胰蛋白酶抑制剂</td><td>沃辛顿</td></tr><tr><td>型胶原酶1 </td><td>沃辛顿</td></tr><tr><td>鼠尾胶原蛋白1型</td><td>屋宇署科学</td></tr><tr><td> FURA-2 </td><td>分子探针</td></tr><tr><td>标用于RT-PCR的第一链合成系统</td><td> Invitrogen公司</td></tr><tr><td>徕卡TCS的SP2光谱共焦显微镜</td><td>莱卡微系统公司</TD&gt; </tr><tr><td>愉-1成像站和MetaMorph软件</td><td>分子器件</td></tr><tr><td>小精尖钳和额外的鸟语花香剪刀</td><td>罚款科学工具</td></tr></tbody></table>

isolation and culture of human fungiform taste papillae cells

味觉细胞是高度专业化的，具有独特的组织学，分子生物学和生理特性，允许一个简单的刺激和食品中含有复杂的化学分子的广泛检测。在人类，个体的菌状乳头，包含从零到多达20个味蕾。有没有人的味觉细胞培养建立的协议，尽管保持文化品位乳头细胞的多种细胞周期的能力将是相当实用，这些独特的感官细胞的特征分子，再生和功能特性。味觉细胞的早先的研究已经完成，使用新鲜细胞原代培养，外植体培养，从啮齿类动物或半完整的味蕾，在组织切片1,2,3,4。虽然这些准备工作的每个人都有优势，将有长远的文化发展提供了重要的好处，特别是对味觉细胞增殖和复合驱研究rentiation。几个团体，包括我们在内，一直在味觉细胞培养模型的建立和发展有兴趣。大多数尝试以文化品位细胞报道有限的活力，细胞通常不超过3-5ð5,6,7,8持久。我们最近报道对啮齿动物的味觉细胞9扩展文化的一个成功的方法。我们在这里报告的第一次孤立的人类蕈状味觉乳头细胞在体外培养体系的建立。活检取自人体的菌状乳头细胞成功地保持了八个多通道（12个月）没有丧失活力的文化。细胞表现出成熟的味觉细胞特征的许多分子和生理功能。 Gustducin和β2，磷脂酶C（PLC - β2）的mRNA在许多细胞，通过逆转录 - 聚合酶链反应检测和测序证实。免疫细胞化学分析表明，在场的阵风ducin和PLC-β2培养味觉细胞中的表达。体外培养的人蕈状细胞也表现出适当的浓度表明，味觉受体和信号通路至少是目前的几种口味刺激细胞内钙离子的增加。这些结果充分表明，成年人类的味觉细胞可以产生和保持为至少8个通道。许多人保留着细胞的急性分离的从成年味道上皮细胞，以支持其作为一种模式味觉系统的使用和生理生化特性。这个系统将使参与增殖，分化和功能在哺乳动物味觉受体细胞， 在体外制备过程的进一步研究。 蕈状人的味觉乳头建立人力蕈状细胞培养的捐赠研究适当的知情同意后，根据研究进行了审查协议由IRB委员会批准。舒尔曼联营机构审查委员会公司，俄亥俄州辛辛那提市批准的协议（＃0934）。书面协议下面是根据报告公布的参数由Ozdener 等 。 2011年10。

Watch this Scientific Journal Video about Isolation and Culture of Human Fungiform Taste Papillae Cells at JoVE.com

Isolation and Culture of Human Fungiform Taste Papillae Cells

我们旨在建立一个孤立和维护人类菌状的味道乳头细胞的长期文化重现协议。活检取自人体的菌状乳头细胞成功地保持了八个多通道（12个月）没有丧失活力的文化。

人力蕈状味觉​​乳头细胞的分离和培养

Taste cells are highly specialized, with unique histological, molecular and physiological characteristics that permit detection of a wide range of simple ...

Research

JoVE Journal

Neuroscience

Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles

从人类毛囊神经嵴干细胞的分离和培养的

Hair follicles undergo lifelong growth and hair cycle is a well-controlled process involving stem cell proliferation and quiescence. Hair bulge is a ...

科研

神经科学

Lectin-based Isolation and Culture of Mouse Embryonic Motoneurons

基于凝集素的分离和培养的小鼠胚胎运动神经元

Spinal motoneurons develop towards postmitotic stages through early embryonic nervous system development and subsequently grow out dendrites and axons. ...

Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay

人类胶质母细胞瘤肿瘤细胞的分离和扩大使用的神经球含量

Stem-like cells have been isolated in tumors such as breast, lung, colon, prostate and brain. A critical issue in all these tumors, especially in ...

Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol

大鼠胚胎神经细胞的分离和培养：一个快速议定书“

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

从小鼠胚胎神经管的神经嵴细胞的分离和培养

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

Isolation and Culture of Mouse Cortical Astrocytes

小鼠脑皮层星形胶质细胞的分离和培养

Astrocytes  are an abundant cell type in the mammalian brain, yet much remains to be  learned about their molecular and functional characteristics. In ...

Isolation and Culture of Endothelial Cells from the Embryonic Forebrain

从胚胎前脑内皮细胞的分离和培养

Embryonic brain endothelial cells can serve as an important tool in the study of angiogenesis and neurovascular development and interactions. The two ...

Denver Papillae Protocol for Objective Analysis of Fungiform Papillae

为菌状乳头的客观分析丹佛乳头协议

The goal of the Denver Papillae Protocol is to use a dichotomous key to define and prioritize the characteristics of fungiform papillae (FP) to ensure ...

Taste Preference Assay for Adult Drosophila

Taste Preference Assay for Adult Drosophila

口味偏好测定成人<em&gt;果蝇</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

Isolation and Culture of Adult Neural Stem Cells from the Mouse Subcallosal Zone

分离和成人神经干细胞的培养从鼠标Subcallosal区

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation ...

人力蕈状味觉​​乳头细胞的分离和培养

人力蕈状味觉乳头细胞的分离和培养