Abstract

The endomembrane system of mammalian cells provides massive capacity for the segregation of biochemical reactions into discrete locations. The individual organelles of the endomembrane system also require the ability to precisely transport material between these compartments in order to maintain cell homeostasis; this process is termed membrane traffic. For several decades, researchers have been systematically identifying and dissecting the molecular machinery that governs membrane trafficking pathways, with the overwhelming majority of these studies being carried out in cultured cells growing as monolayers. In recent years, a number of methodological innovations have provided the opportunity for cultured cells to be grown as 3-dimensional (3D) assemblies, for example as spheroids and organoids. These structures have the potential to better replicate the cellular environment found in tissues, and present an exciting new opportunity for the study of cell function. In this mini-review we summarise the main methods used to generate 3D cell models, and highlight emerging studies that have started to use these models to study basic cellular processes. We also describe a number of pieces of work that potentially provide the basis for adaptation for deeper study of how membrane traffic is coordinated in multicellular assemblies. Finally, we comment on some of the technological challenges that still need to be overcome if 3D cell biology is to become a mainstream tool towards deepening our understanding of the endomembrane system in mammalian cells.

Highlights

  • The cell can be defined as the basic building block of the tissues and organs that build the human body

  • Internalised material can be directed into the endosomal-lysosomal system, or in some cases it can be trafficked towards the Golgi apparatus and the endoplasmic reticulum (ER) via the retrograde pathway

  • These pathways provide a convenient mechanism for the cell to be able to connect all the key compartments of the endomembrane system, they facilitate the recycling of trafficking machinery molecules back to their site of origin, and importantly they maintain the overall homeostasis of the cell

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Summary

Introduction

The cell can be defined as the basic building block of the tissues and organs that build the human body. Optical microscopy has become a mainstay technique for study of membrane trafficking pathways, but issues such as light scattering present particular challenges when imaging living 3D models [25]. While this spheroid study was small-scale, it again points to the fact that there are differences in the mechanics of the endomembrane system depending on the growth format of the cells.

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