Science – FluoSphera

Years of clinical trials to know the behavior of a single drug before FDA approval.

Of drugs are metabolized by the liver, affecting their effects on other tissues.

Of clinical failure because drug activity is weakly predicted by preclinical studies.

High-Throughput methods with high physiological relevance

Methods able to mimic communications between distant tissues

Human-based in vitro methods

Most molecules in contact with human beings cannot be characterized in patients. In vitro methods mimicking the complexity of human physiology are needed for a large-scale prediction of the effects of most molecules with a good reliability.

Multi-culture methods with several human tissues have to be developed to recapitulate the physiological exchanges of molecules between organs, their metabolic transformation, and their tissue-specific effects.

Human-based in vitro methods are needed to overcome the strong inter-species variabilities observed between animal models and humans.

Our Solution

The properties of light were used for the most important technological breakthroughs, like laser, optical fiber, or next-generation sequencing. FluoSphera uses the precision of fluorescent light to simultaneously measure multiple biological processes in multiple human cell types in co-culture. Our multi-culture assays mimic the communications between human organs, for a reliable prediction of the effects of compounds on human health, unlike current mono-culture assays.

Technology

Fluorescent Hollow Capsule

Fluorescent Spheroid

High-Content Screening

Recognition of encapsulated cell types with permeable and color-coded capsules in co-culture (up to 5 different tissues).

Detection of biological processes in encapsulated spheroids with fluorescent reporter genes and/or fluorescent reagents.

Acquisition and analysis of fluorescent patterns with a strong multiplexing capacity with the technology of High-Content Screening (HCS).

Cell Encapsulation with Microfluidics

Image acquisition with HCS

A unique combination

Compartmentalization & Permeability

3D Cell Culture

Spheroid Size Control

The capsule physically separates the different tissues from each other and allows them to exchange soluble factors, like in human body.

Within a few days after encapsulation, cells form spheroids, well-known to recapitulate the spatial organization of tissues.

The capsule controls the size of the spheroid below 300 µm, compatible with oxygen and nutrients diffusion, like in perfused tissues.

Non-Invasive Tissue Identification

A capsule color is assigned to each tissue type for their recognition even in a complex multi-culture, without labelling cells.

Multiplexing in Real-Time

The combination of fluorescent intensities of cells with the fluorescent capsules allows for the simultaneous measurement of multiple activities in multiple tissues in real-time.

Compatibility with HCS

The fluorescent patterns generated by FluoSphera assays are perfectly adapted to HCS, one of the most powerful imaging technology in Life Sciences.

Example of Applications

Multiplexed measurement in co-culture

Image acquisition of a BC/LC co-culture of encapsulated cells

Co-culture of breast cancer (BC) cells in far-red capsules (blue) with liver cancer (LC) cells in unlabeled capsules (non-fluorescent). Both cell types stably express the proliferation reporter FUCCI (proliferating cells in green, latent cells in red). The cell segmentation profile obtained with the HCS technology is shown at the right. It was used to quantify the different fluorescent intensities in each type of capsule.

Simultaneous measurement of two proliferation states in BC cells and in LC cells in co-culture.

Recapitulation of distant physiological communications to better predict the effect of molecules

Inhibition of ERα in BC cells in a BC/LC co-culture

Breast cancer (BC) cells were co-cultured or not with liver cancer (LC) cells and were treated with increasing concentrations of Tamoxifen (Tam) or 4-hydroxytamoxifen (4-OHT, an active metabolite of Tam) as control. The activity of Estrogen Receptor α (ERα) was measured in BC cells.

Tam is an anticancer drug against breast cancer. Once in the human body, Tam is first metabolized by the liver to an active metabolite then transported to the breast tumor to inhibit ERα and to stop proliferation. We observe on the graph that the efficiency of Tam is increased by almost 20-fold in the BC/LC co-culture when compared to the BC mono-culture. This shows that with FluoSphera assays we can mimic physiological communications between organs and that it is essential to accurately predict the effects of molecules on human health (here the efficiency of Tam to treat breast cancer).

Multiplexed measurement in co-culture

Recapitulation of distant physiological communications to better predict the effect of molecules

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