SPHEROID CELL CULTURE using Microfluidics 

Spheroids are three-dimensional self-assembly of cells that better reproduce cell-to-cell interactions and can mimic in vivo environments more [1]. They can be formed using one or multiple cell types depending on the organ studied. They are considered the best tumor cellular models [2] and can also be used to study neurodegenerative diseases [3].

3D cell culture models better predict drug efficacy compared to animal models, and they tend to replace them in the pre-clinical testing phase (“3R”: Replacement, Reduction, Refinement)

Spheroid formation process in a microwell-based µSFC: (A) Introduction of a cell suspension to the chip inlet. The cell suspension fills all the microchannels and microwells rapidly due to the capillary effect; (B) Cells start depositing on the bottom of the microchannels and microwells; (C) Pure culture medium flows through the chip to rinse the excess cells without disturbing the cells lying on the microwell bottom; (D) Cell secretions and signaling lead to the establishment of cell-cell interactions on the non-adherent microwell bottom; (E) Driving spheroid formation under a perfusing flow of culture medium [4].

–> Learn more about the advantages of 3D cell culture in our introduction to 3D cell culture.

Culturing spheroids in microfluidics brings critical advantages compared to classical methods:

• Lower experiment costs by reducing the amount of reagent used
• Reduced number of cells required for each experiment
• Cultured spheroids experience more physiological shear stress
• A single spheroid can be cultivated and observed simultaneously
• Even long experiments can be easily automated and don’t require human intervention
• Improved oxygen and nutrition supply for cells
• Better reproducibility and uniformity
• Easily inject precise volumes of different drugs or compounds

Thus, microfluidics is the best solution for performing spheroid culture and drug screening.

Elveflow instruments are specially designed for this kind of application due to their stability, user-friendliness, accuracy, and flow control.  Plus, Elveflow provides the best flow control on the market.

Spheroid formation in microfluidic wells: (1) cell seeding, (2) aggregation within first 24 h, (3) medium flush and (4) compact spheroid formation within next 24 h. Scale bars correspond to 50 mm. Ziółkowska et al. [5]

1. Astashkina, Anna, Brenda Mann, and David W. Grainger. “A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity.” Pharmacology & therapeutics 134.1 (2012): 82-106.
2. Friedrich, Juergen, Reinhard Ebner, and Leoni A. Kunz-Schughart. “Experimental anti-tumor therapy in 3-D: spheroids–old hat or new challenge?.” International journal of radiation biology 83.11-12 (2007): 849-871.
3. Słońska, Anna, and Joanna Cymerys. “Application of three-dimensional neuronal cell cultures in the studies of mechanisms of neurodegenerative diseases.” Postepy Higieny i Medycyny Doswiadczalnej (Online) 71 (2017): 510-519.
4. Moshksayan, Khashayar, et al. “Spheroids-on-a-chip: Recent advances and design considerations in microfluidic platforms for spheroid formation and culture.” Sensors and Actuators B: Chemical 263 (2018): 151-176.
5. Karina Ziółkowska; Agnieszka Stelmachowska; Radosław Kwapiszewski; Michał Chudy; Artur Dybko; Zbigniew Brzózka (2013). Long-term three-dimensional cell culture and anticancer drug activity evaluation in a microfluidic chip. , 40(1)