High-Resolution Microscopic Characterization of Tunneling Nanotubes in Living U87 MG and LN229 Glioblastoma Cells

Sep 21, 2026

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Tunneling nanotubes (TNTs) are fine nanometer-scale membrane connections enabling communication between distant cells and shaping cellular organization. Their involvement in cancer biology is proposed to influence aggressive behavior and therapeutic resistance, particularly in glioblastoma. This study characterizes TNT properties in U87 MG and LN229 glioblastoma lines using high-resolution live-cell STED microscopy for diameter and live-cell confocal microscopy for length, morphology, lifetime, and formation, evaluating membrane labeling approaches and recommending live-cell study designs.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages27

Reading Time1 h 8 min

AccessFree to download

Abstract

Keywords

Introduction

Cellular communication and cancer challenges

TNT mechanisms and discovery

TNT diversity and formation theories

cells Article High-Resolution Microscopic Characterization of Tunneling Nanotubes in Living U87 MG and LN229 Glioblastoma Cells Nicole Matejka *, Asieh Amarlou, Jessica Neubauer, Sarah Rudigkeit and Judith Reindl Citation: Matejka, N.; Amarlou, A.; Neubauer, J.; Rudigkeit, S.; Reindl, J. High-Resolution Microscopic Characterization of Tunneling Nanotubes in Living U87 MG and LN229 Glioblastoma Cells. Cells 2024, 13, 464. [https://doi.org/10.3390/](https://doi.org/10.3390/)[ ](https://doi.org/10.3390/)cells13050464 Academic Editors: Gregor Drummen and Hellen Ishikawa-Ankerhold Received: 2 February 2024 Revised: 4 March 2024 Accepted: 5 March 2024 Published: 6 March 2024 Copyright: © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) . Institute for Applied Physics and Measurement Technology, University of the Bundeswehr Munich, 85577 Neubiberg, Germany; [asieh.amarlou@tum.de](asieh.amarlou@tum.de) (A.A.); [jessica.neubauer@unibw.de](jessica.neubauer@unibw.de) (J.N.); [sarah.rudigkeit@unibw.de](sarah.rudigkeit@unibw.de) (S.R.); [judith.reindl@unibw.de](judith.reindl@unibw.de) (J.R.) * Correspondence: nicole.matejka@unibw.de Abstract: Tunneling nanotubes (TNTs) are fine, nanometer-sized membrane connections between distant cells that provide an efficient communication tool for cellular organization. TNTs are thought to play a critical role in cellular behavior, particularly in cancer cells. The treatment of aggressive cancers such as glioblastoma remains challenging due to their high potential for developing therapy resistance, high infiltration rates, uncontrolled cell growth, and other aggressive features. A better understanding of the cellular organization via cellular communication through TNTs could help to find new therapeutic approaches. In this study, we investigate the properties of TNTs in two glioblastoma cell lines, U87 MG and LN229, including measurements of their diameter by high-resolution live-cell stimulated emission depletion (STED) microscopy and an analysis of their length, morphology, lifetime, and formation by live-cell confocal microscopy. In addition, we discuss how these fine compounds can ideally be studied microscopically. In particular, we show which membrane-labeling method is suitable for studying TNTs in glioblastoma cells and demonstrate that live-cell studies should be preferred to explore the role of TNTs in cellular behavior. Our observations on TNT formation in glioblastoma cells suggest that TNTs could be involved in cell migration and serve as guidance. Keywords: tunneling nanotube; glioblastoma; cellular communication; cancer treatment; live-cell microscopy; high-resolution microscopy 1. Introduction The ability of cells to rapidly adapt and reorganize themselves when their environment and living conditions change has been an important aspect of global research interest since the beginning of cell research. Efficient cellular communication is a key driver in wound healing, immune response, embryonic development, and more, as well as in disease control and prevention. One of medicine’s greatest challenges is the fight against cancer. According to the International Agency for Research on Cancer’s (IARC) World Cancer Report 2020, 29.8% of premature deaths from noncommunicable diseases (NCDs) worldwide in 2016 were due to cancer, making it the leading cause of death worldwide [1] . Therefore, the role of cellular communication in cancer treatment has been intensively studied to better understand the complexities involved in the interaction of different cell types in the tumor microenvironment, cancer invasion, and progression, and to identify novel therapeutic strategies [2

FAQs

What are tunneling nanotubes (TNTs) and why are they important in cancer?

TNTs are nanometer-sized membrane connections that allow efficient communication between distant cells. They are thought to influence cellular behavior, including in cancer cells such as glioblastoma.

How were TNT properties measured in this study?

TNT diameter was measured using high-resolution live-cell STED microscopy. Length, morphology, lifetime, and formation were analyzed by live-cell confocal microscopy.

Which microscopy approach is recommended for studying TNTs in glioblastoma cells?

The study demonstrates that membrane-labeling method choice matters and shows that live-cell studies should be preferred to explore TNT roles in cellular behavior.

Sep 21, 2026

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