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Cell spheroids preparation: A novel low-cost technique

Kevin Bellofatto, Switzerland

Research assistent
Surgery. Transplantation.
Hospitaux Universitaires de Geneve

Abstract

Cell spheroids preparation: A novel low-cost technique

Ibrahim Fathi1, Nouran Zahran1, Kevin Bellofatto2, Hagar Elkafrawy1, Radwa Mehanna1.

1Center of Excellence for Research in Regenerative Medicine and Applications (CERRMA), Faculty of Medicine, Alexandria University, Alexandria, Egypt; 2Cell Isolation and Transplantation Centre, Department of Surgery, Geneva University Hospitals, Geneva, Switzerland

Introduction. Several methods have been developed to produce cell spheroids in the laboratory; including rocking method, agarose silicone moulds, and micro-well plates. However, most of available ready-made silicone moulds and micro-well plates are considered as expensive options. We here describe a simple low-cost method to produce agarose micro-wells using commercially available Japanese floral frogs. Methods. 2500 µL of sterile 1.5% w/v agarose were placed per well in 12-well plates and commercially available stainless steel Japanese floral frogs (autoclaved) were placed in the well-plates upside down to produce the micro-grooves inside the agarose. After gelation, the floral frogs were removed, and the micro-wells were used for cell seeding (Figure 1). 20,000 HepG2 cells/well were used for demonstration. Results: HepG2 spheroids were produced on day 3-5 after cell seeding and were retrieved through gentle pipetting. The MTT assay will be performed in comparison to spheroids made in ready-made spheroids microplates and 2D culture and the results will be presented at the conference. Conclusion. Using Japanese floral frogs is a simple and low-cost method to produce spheroids. The testing of cell viability and proliferation in the new model is still ongoing. 

 

References:

[1] Spheroids
[2] Agarose
[3] HepG2

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