arXiv subjects

Simone Scalise

Publications and source records attributed to Simone Scalise.

4 recordsLinked to original sources

arXiv2025

A mother-machine microfluidic device for non-adherent mammalian cells reveals the population growth strategies

We develop a mother machine-like microfluidic device specifically designed to track the proliferation of T-cells via live-cell microscopy. Although numerous microfluidic setups have been developed to study cell proliferation at the single-cell level, most of them are optimized for use on adherent cells. Here, we present a device to track the proliferation of suspension cells, featuring an array of microchannels that trap cells, easing their monitoring while allowing for controlled growth conditions. Each microchannel, whose geometry has been optimized through computational fluid dynamics simulations, allows a single cell to enter and proliferate while maintaining a continuous flow of nutrients, ensuring long-term monitoring over multiple generations. We show the advantages of this system in characterizing the proliferation of human leukemia T-cells. In particular, we follow the growth and division over multiple generations, finding that cells exhibit a slightly asymmetric volume division where deviations in the size are compensated by a size-like division strategy. Overall, our device design can be easily adapted and used to study different cell types and sizes while maintaining the same high trapping efficiency.

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arXiv2024

Robust assessment of asymmetric division in colon cancer cells

Asymmetric partition of fate determinants during cell division is a hallmark of cell differentiation. Recent work suggested that such a mechanism is hijacked by cancer cells to increase both their phenotypic heterogeneity and plasticity and in turn their fitness. To quantify fluctuations in the partitioning of cellular elements, imaging-based approaches are used, whose accuracy is limited by the difficulty of detecting cell divisions. Our work addresses this gap proposing a general method based on high-throughput flow cytometry measurements coupled with a theoretical framework. We applied our method to a panel of both normal and cancerous human colon cells, showing that different kinds of colon adenocarcinoma cells display very distinct extents of fluctuations in their cytoplasm partition, explained by an asymmetric division of their size. To test the accuracy of our population-level protocol, we directly measure the inherited fractions of cellular elements from extensive time-lapses of live-cell laser scanning microscopy, finding excellent agreement across the cell types. Ultimately, our flow cytometry-based method promises to be accurate and easily applicable to a wide range of biological systems where the quantification of partition fluctuations would help accounting for the observed phenotypic heterogeneity and plasticity

q-bio.CB↗

arXiv2024

Probing leukemia cells behavior under starvation

The ability of a cancer cell population to achieve heterogeneity in their phenotype distributions offers advantages in tumor invasiveness and drug resistance. Studying the mechanisms behind such observed heterogeneity in mammalian cells presents challenges due for instance to the prolonged proliferation times compared to widely studied unicellular organisms like bacteria and yeast. Here, we studied the response of leukemia cell populations to serum starvation via a protocol, we recently developed, that makes use of live cell fluorescence and flow cytometry in combination with a quantitative analytical model to follow the population proliferation while monitoring the dynamics of its phenotype distributions. We found that upon switching between a serum-rich to a serum-poor media, leukemia cells (i) maintain a memory of the previous environment up to one generation even in the presence of severe medium-depletion, before (ii) adapting their growth and division rates to the novel environment while preserving a sizer-like division strategy. Finally, looking at the mitochondria content of the proliferating vs non-proliferating cells, we found that the latter is characterized by a higher number of older mitochondria, suggesting a possible functional role of the observed asymmetric partitioning of (aged) mitochondria in leukemia cells.

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arXiv2023

Determining cancer cells division strategy

Heterogeneity in the size distribution of cancer cell populations has been recently linked to drug resistance and invasiveness. However, despite many progresses have been made in understanding how such heterogeneous size distributions arise in fast-proliferating cell types -like bacteria and yeast-, comprehensive investigations on cancer cell populations are still lacking mainly due to the difficulties of monitoring the proliferation of the time scales typical of mammalian cells. From a reductionist cell dynamics point of view, the strategies allowing size homeostasis are roughly grouped into three classes, \emph{i.e.} timer, sizer, or adder. These strategies are empirically distinguishable given the phenomenological measurable relationship between the cell size at birth and at division, which requires following the proliferation at the single-cell level. Here, we show how it is possible to infer the growth regime and division strategy of leukemia cell populations using live cell fluorescence labeling and flow cytometry in combination with a quantitative analytical model where both cell growth and division rates depend on powers of the cell size. Using our novel approach, we found that the dynamics of the size distribution of leukemia Jurkat T-cells is quantitatively reproduced by (i) a sizer-like division strategy, with (ii) division times following an Erlang distribution given by the sum of at least three independent exponentially-distributed times and (iii) fluctuations up to 15\% of the inherited fraction of size at division with respect to the mother cell size. Finally, we note that our experimental and theoretical apparatus can be easily extended to other cell types and environmental conditions, allowing for a comprehensive characterization of the growth and division model different cells can adopt.

q-bio.CB↗