Cell3: a new vision for study of the endomembrane system in mammalian cells.

Margaritha M Mysior,Jeremy C Simpson

doi:10.1042/bsr20210850c

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 summarize 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 toward 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
Micropatterning on glass-bottomed multi-well plates opens up the potential for using confocal high-content screening microscopy to image these spheroids with subcellular resolution, revealing the morphology of organelles such as the Golgi apparatus in each of the cells in the spheroid (Figure 2B)
This in turn allows cell-based assays to be carried out, for example treatment of the spheroids with classical endomembrane perturbation drugs, such as brefeldin A (BFA), which have proved to be highly useful tools for our deepening our understanding of the endomembrane system [55]

Summary

Introduction

The cell can be defined as the basic building block of the tissues and organs that build the human body. One significant challenge of working with 3D cell models is selecting a growth method that finds the best compromise between providing a physiologically relevant assembly, and which is amenable to their study, for example using high-resolution imaging. Image analysis included volumetric measurements of the entire spheroid and the individual nuclei within each cell, providing a high level of detail with respect to the changes seen on

Results

Conclusion