Microfluidic Platform for Efficient Microtissue Generation
The generation of organoids and spheroids remains a highly labor-intensive and technically demanding process, often requiring meticulous manual handling and extended optimization to achieve reliable results. Despite significant interest in 3D culture models, the real-world success rate for consistent, viable microtissue formation is relatively low, particularly when working with primary human cells. As a result, many researchers are forced to spend substantial time troubleshooting cell aggregation and viability issues rather than advancing translational research goals. This challenge is further amplified by the growing demand for physiologically relevant, human-based models that better replicate in vivo conditions—highlighting the need for more efficient, automated solutions that can bridge the gap between basic research and clinically meaningful outcomes.
To overcome these limitations and accelerate the adoption of reliable human-based models, we embarked on the development of a microfluidic-based platform specifically designed to automate and standardize spheroid and organoid generation. Our goal was to eliminate the bottlenecks of manual workflows, reduce variability, and significantly improve the success rate of microtissue formation—even with challenging primary human cells. By integrating precise fluid control, optimized microenvironment conditions, and automation-friendly design, our platform enables researchers to focus on meaningful translational studies rather than repetitive and error-prone cell culture procedures.
Microbead Generation
The microfluidic process begins by combining cells and extracellular matrix (ECM) hydrogel to form a dispersed phase, which is then processed within a microfluidic chip. ECM hydrogels in most cases are temperature-sensitive, allowing them to be handled as a liquid at 4°C and then transformed into a gel upon heating. By employing the single emulsion method, with fluorinated oil (HFE) serving as the continuous phase, we achieve a high-throughput production of approximately 1,000 microbeads per minute. The addition of a fluorinated surfactant stabilizes the droplet emulsion in HFE, ensuring the integrity of droplets during subsequent procedures.
The flow rates of the dispersed and continuous phases are determined by the applied pressure, channel geometry, and fluid properties.
Our microfluidic design incorporates magnetic stirring
within the dispersed phase during droplet generation.
This mechanism ensures a uniform distribution of cells
within each microbead.
without magnetic stirring
with magnetic stirring
Matrigel Polymerization
After collecting the desired quantity of droplets, they are incubated at a physiological temperature to induce the polymerization of the ECM-based hydrogel. Optimizing the concentration of ECM when mixed with cell suspension is crucial for achieving robust microbeads with enhanced mechanical stability.
ECM protein concentration (Corning)
4.5 mg/ml
3.6 mg/ml
3 mg/ml
2.6 mg/ml
The recovery of microbeads from oil is efficiently achieved using an oleophilic absorbent, such as a polyvinylidene fluoride (PVDF) plate made by non-solvent induced phase separation fabrication. This method is highly effective due to the ability of the porous plate to selectively absorb oil while preserving the integrity of the microbeads.
Following oil absorption, the microbeads are transferred into cell medium, providing the necessary nutrients to support cell growth and facilitate Microtissue formation. While approximately 30% of microbeads are lost during the adsorption process, this method still yields a substantial number—typically around 10,000 microbeads per procedure when using 200μL dispersed phase.
Incubation for Microtissue Formation
After the microbead suspension is dispensed into well plates it is incubated under controlled conditions: 37°C and 5% CO2. This environment supports the growth and aggregation of cells into Microtissues. The Microtissue formation timeline varies significantly depending on the cell type. Immortalized cell lines typically form Microtissues within 24 hours, reflecting their rapid growth and aggregation characteristics.
MDA-MB231 (50 cells/ml)
Microbeads suspension in oil
Recovered from oil
24h of incubation
In contrast, to immortalized cells primary cells may require up to three weeks for Microtissue formation, as they often exhibit slower growth rates and need more time to establish stable cell-cell interactions. For primary cells aggregation and Microtissue formation more depends on cell concentration and time of incubation.
HCC cells. Donor 1 (80 cells/microbead)
Day 1
48 hours
7 days
HCC cells. Donor 2 (40 cells/microbead)
Day 1
2 weeks
3 weeks
Hepatocellular carcinoma (HCC)-derived spheroid formation is notoriously challenging, with conventional methods rarely achieving acceptable success rates for reliable disease modeling and drug screening. The low viability and poor aggregation efficiency of primary HCC cells often result in inconsistent and non-reproducible models, limiting their translational relevance. The SpheroFlow™ platform overcomes these barriers, delivering a remarkable 90% success ratein generating uniform, viable HCC-derived spheroids. To further streamline research efforts, we offer a curated library of SpheroFlow-prequalified HCC cells and other primary and cancer cell types, ensuring immediate compatibility with the system and reliable outcomes from the very first experiment.
Untreated control - HCC spheroids
100 micromole/L of carboplatin
At Preci, we offer an advanced procedure for utilizing microfluidic technology in combination with extracellular matrices and cells to automate the generation and analysis of tumor Microtissues. Our solutions are designed to bridge the gap between traditional cell culture techniques and clinical trials, providing a more reliable and efficient pathway for understanding cancer biology and evaluating therapeutic strategies.
1000 micromole/L of carboplatin
