Mammalian Cell Aggregates
Mammalian Cell Aggregates - Related Videos
08:59
4D Imaging of Protein Aggregation in Live Cells
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Cited by 4 •
2013
Cellular viability depends on timely and efficient management of protein misfolding. Here we describe a method for visualizing the different potential fates of a misfolded protein: refolding, degradation, or sequestration in inclusions. We demonstrate the use of a folding sensor, Ubc9ts, for monitoring proteostasis and aggregation quality control in live cells using 4D microscopy.
04:12
Protein Aggregate Formation Assay: A Method to Detect and Quantify Protein Aggregation in Cultured Cells upon Induction by Proteasome Inhibitor
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2025
In this video, we demonstrate a cell-based protein aggregation assay using proteasome inhibitors, which block proteasome activity, preventing misfolded, mutant proteins, fused to a fluorescent label, from undergoing ubiquitin-dependent proteasomal degradation, leading to their accumulation within the cell cytoplasm. The protein aggregates are then visualized and quantified by fluorescence microscopy.
05:40
An Orbital Shaking Culture of Mammalian Cells in O-shaped Vessels to Produce Uniform Aggregates
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Cited by 6 •
2019
Here we present a protocol for using O-shaped vessels, specialized for suspension cultures of cellular aggregates, with orbital shaking. The HEK293 cells grown in this bag form more homogeneous aggregates than those grown in conventional culture vessels.
08:39
Electroporation of Functional Bacterial Effectors into Mammalian Cells
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Cited by 7 •
2015
Electroporation was used to insert purified bacterial virulence effector proteins directly into living eukaryotic cells. Protein localization was monitored by confocal immunofluorescence microscopy. This method allows for studies on trafficking, function, and protein-protein interactions using active exogenous proteins, avoiding the need for heterologous expression in eukaryotic cells.
09:43
Measurement of Heme Synthesis Levels in Mammalian Cells
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Cited by 12 •
2015
Altered intracellular heme levels are associated with common diseases such as cancer. Thus, there is a need to measure heme biosynthesis levels in diverse cells. The goal of this protocol is to provide a fast and sensitive method to measure and compare the levels of heme synthesis in different cells.
FAQs
Aggregate size shapes internal oxygen and nutrient gradients, which can alter cell viability, differentiation, and signaling. These gradients create spatial differences within the same three-dimensional structure rather than exposing every cell to identical conditions. Controlling size therefore helps bioengineers investigate how local environments influence tissue-like behavior and model the organization of developing or diseased tissue.
Cell-cell adhesion brings neighboring cells into a coordinated structure, while extracellular matrix remodeling helps reshape and stabilize the developing aggregate. Together, these processes influence compaction and tissue architecture. Their activity is important because aggregate organization depends not only on how closely cells are positioned, but also on how cells collectively modify their surrounding environment.
Different cellular compositions can produce distinct patterns of signaling and differentiation within an aggregate. At the same time, internal oxygen and nutrient gradients expose cells to different local conditions. The combination of cell identity, spatial position, and available resources allows aggregates to model coordinated tissue behavior and helps researchers examine how microenvironments influence cell fate.
Flat-surface cultures provide a comparatively simple two-dimensional setting, whereas aggregates allow cells to organize through close contact and collective growth in three dimensions. This spatial arrangement better represents aspects of tissue architecture, including internal gradients and extracellular matrix remodeling. The comparison helps researchers select aggregates when cell organization and tissue-like interactions are central to the experiment.
Formation begins by bringing mammalian cells into close contact under conditions that limit their attachment to a flat surface. Cells can then adhere to one another, compact collectively, and remodel extracellular matrix as the structure develops. Researchers can evaluate the resulting aggregates by considering their size, cellular composition, organization, and effects on viability or differentiation.
These aggregates are useful when researchers need a simplified three-dimensional model for tissue development, disease processes, or therapeutic-response studies. They can also support organoid-like modeling, where organized cell behavior provides information beyond a flat culture. Their value comes from connecting cellular interactions and spatial gradients with measurable changes in tissue-like function or response.
In tissue engineering, aggregates provide building units in which coordinated cell behavior can support functional tissue formation. Their organization, composition, and matrix remodeling can be examined as part of a larger engineered system. This approach helps researchers study how cells assemble, communicate, and develop tissue-like architecture while accounting for internal oxygen and nutrient conditions.