Identification and Characterization of Tunneling Nanotubes for Intercellular Trafficking.
Identification and Characterization of Tunneling Nanotubes for Intercellular Trafficking.
复制标题
DOI：
10.1002/0471143030.cb1210s67
复制
发表时间：
2015-06-01
影响因子：
--
通讯作者：
Zurzolo, Chiara
中科院分区：
文献类型：
--
作者：
Abounit, Saida;Delage, Elise;Zurzolo, Chiara
Tunnel nanotubes（TNT）是一种薄膜通道，在远端细胞之间提供直接的细胞质连接。它们通常在不同的细胞培养物中观察到，越来越多的证据支持它们在细胞间通讯和病原体转移中的作用。然而，对TNT的研究存在几个陷阱（例如，难以保存这种精细的结构，可能与其他突起混淆，结构和功能异质性等）因此需要彻底设计的方法。本单元中描述的方法代表了细胞培养中TNT（或TNT样结构）表征的指南。具体地，提供了优化的方案，以（1）鉴定TNT和它们内部存在的细胞骨架元件;（2）评估细胞培养物中的TNT频率;（3）明确地将它们与其他细胞连接或突起区分开;以及（4）监测它们在活细胞中的形成。最后，本单元描述了如何评估TNT介导的细胞成分的细胞间转移，这是识别功能性TNT的基本标准。
Tunneling nanotubes (TNTs) are thin membranous channels providing direct cytoplasmic connection between remote cells. They are commonly observed in different cell cultures and increasing evidence supports their role in intercellular communication and pathogen transfer. However, the study of TNTs presents several pitfalls (e.g., difficulty in preserving such delicate structures, possible confusion with other protrusions, structural and functional heterogeneity, etc.) and therefore requires thoroughly designed approaches. The methods described in this unit represent a guideline for the characterization of TNTs (or TNT-like structures) in cell culture. Specifically, optimized protocols to (1) identify TNTs and the cytoskeletal elements present inside them; (2) evaluate TNT frequency in cell culture; (3) unambiguously distinguish them from other cellular connections or protrusions; and (4) monitor their formation in living cells are provided. Finally, this unit describes how to assess TNT-mediated cell-to-cell transfer of cellular components, which is a fundamental criterion for identifying functional TNTs.