As 3D cell culture is a recent development, it is not always easy to identify which tools to use. Here are three of them, ranging from the growing of compact and reproducible spheroids, to their study under confinement.
Principle Stampwell U-Shape is a stamp designed to easily create U-bottom wells in any type of hydrogels such as agarose, alginate, phytagel, methycellulose, etc. This forms an array of non-adherent wells in which cell suspension can be loaded to watch cell aggregate in individual 3D cell structures. The same stamp with flat bottom wells, called Stampwell V-shape, has been designed to be able to better image the spheroids.
Key Features
Samples immobilized on a similar focal plane
Compatible with a wide variety of hydrogels
Reusable
Cost-Saving compared to low-adherent well-plates such as ULA
Developed by Gaëlle Recher and Camille Douillet (CNRS – Université de Bordeaux, France) Alessandri K, Andrique L, Feyeux M, Bikfalvi A, Nassoy P, Recher G. All-in-one 3D printed microscopy chamber for multidimensional imaging, the UniverSlide. Sci Rep. 2017 Feb 10;7:42378. doi: 10.1038/srep42378. PMID: 28186188; PMCID: PMC5301227.
Growing uniformly-sized spheroids with SpheroTribe
Principle SpheroTribe is a concentrated polymer-based solution that increases the medium viscosity, therefore favouring cell-cell contacts. It offers a simple method to generate homogeneous 3D cell structures with increased control over their shape, minimizing variability between samples and improving the consistency of your downstream experiments such as migration/invasion assays, immunostaining, drug screening or in vivo implantation experiments. SpheroTribe is particularly useful to boost aggregation when working with challenging cells.
Key features • Cell aggregation booster • Increased homogeneity • Easy to use : simply dilute it in your culture media • No need to work on ice
Developed by Guyon, J., Andrique, L., Pujol, N., Røsland, G. V., Recher, G., Bikfalvi, A., & Daubon, T.
AgarSqueezer to mimic the compression of a real cell environment
AgarSqueezer is an innovative cell culture system to grow and image cells under different confinement levels within a physiological rigidity range (1-150 kPa). The confinement system is based on molded agarose pillars, which enables an instant & homogeneous compression on cells, to study their response to short and long-term confinement up to 10 days.
Key features • Homogeneous cell confinement • Tunable pressure in a physiological range : 1-150 kPa • Physiological rigidity • Open access to the reservoir: Possibility to add drugs, and reagents. Easy medium renewal • Long term confinement • Fully compatible with multiple microscopy techniques • Easy to recover coverslip with cells for subsequent molecular analysis: FACS, qPCR, Western-Blot, Immunofluorescence (possible in situ)
Developed by Audrey Prunet, Gilles Simon, Hélène Delanoë-Ayari, Véronique Maguer-Satta and Charlotte Rivière. Prunet, A., Lefort, S., Delanoë-Ayari, H., Laperrousaz, B., Simon, G., Barentin, C., Saci, S., Argoul, F., Guyot, B., Rieu, J. P., Gobert, S., Maguer-Satta, V., & Rivière, C. (2020). A new agarose-based microsystem to investigate cell response to prolonged confinement. Lab on a chip, 20(21), 4016–4030. https://doi.org/10.1039/d0lc00732c
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