Abstract
Gathering precise information on mass density, size and weight of cells or cell aggregates, is crucial for applications in many biomedical fields with a specific focus on cancer research. Although few technical solutions have been presented for single-cell analysis, literature does not cover this aspect for 3D models such as spheroids. Since the research interest on such samples is notably rising, here we describe a flow-apparatus, and the associated physical method and operative protocol for the accurate measurements of mass density, size and weight. The technique is based on the detection of the terminal velocity of a free-falling sample into a specifically conceived analysis flow-channel. Moreover, in order to demonstrate the accuracy and precision of the presented flow-device, analyses were initially carried out on standardized polystyrene beads. Finally, to display the application of the proposed system for biological samples, mass density, size and weight of live SW620 tumor spheroids were analyzed. The combined measurements of such parameters can represent a step toward a deeper understanding of 3D culture models.
Highlights
The past decades have been characterized by the development of three-dimensional (3D) cell culture systems, as they provide more predictive in vitro models to study fundamental cell biology, disease pathophysiology, and the identification of novel therapeutic agents [1,2,3,4,5].the reproducibility of 3D models, in terms of size, shape and 3D architecture, can be difficult to achieve, and their generation is often time- and resource-consuming
This is the first time that a quantitative measurement of mass density, and the simultaneous detection of size and weight, is performed for spheroids
Mass density is generally correlated with compactness, which is qualitatively measured via imaging techniques [36], mathematical models [37], or 3D reconstruction based on bi-dimensional images [13]
Summary
The past decades have been characterized by the development of three-dimensional (3D) cell culture systems, as they provide more predictive in vitro models to study fundamental cell biology, disease pathophysiology, and the identification of novel therapeutic agents [1,2,3,4,5].the reproducibility of 3D models, in terms of size, shape and 3D architecture, can be difficult to achieve, and their generation is often time- and resource-consuming. When monitoring spheroids growth via imaging analyses, attention is focused on their size- and shape-variation over time. This gives indirect information on spheroid compactness, which is often highlighted by a transition from a loose to a tighter cell aggregate [13]. This approach is not fully representative of 3D models, as spheroids undergo specific compaction processes over time, depending on factors such as cellular heterogeneity, growth environment and pharmacological treatments [16,17,18,19]. Aggregates with similar dimensions can have various compositions, related to the metabolic/proliferative cell state, and the extracellular matrix deposition
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