HIGH-THROUGHPUT GENERATION OF HYDROGEL MICRODROPLETS FOR MICROTISSUE ENGINEERING APPLICATION

Objectives: Droplet microfluidics-based strategies are gaining popularity recently for various applications in biotechnology and therapeutics. However, achieving high throughput in producing cell-laden microbeads is still a challenge. Our research shows a reliable, single-run approach for the high-t...

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Bibliographic Details
Published in:International journal of artificial organs 2023-07, Vol.46 (7), p.411
Main Authors: Mukherjee, D, Rudzinska–Radecka, M, Klak, M, Wszola, M, Guzowski, J
Format: Article
Language:English
Subjects:
Biotechnology
Cancer
Cell culture
Cell viability
Cores
Crosslinking
Crosstalk
Droplets
Emulsions
Encapsulation
Fibroblasts
Hydrogels
Microfluidics
Microspheres
Nanoparticles
Organoids
Three dimensional models
Tissue engineering
Tumor microenvironment
Tumors
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Summary:Objectives: Droplet microfluidics-based strategies are gaining popularity recently for various applications in biotechnology and therapeutics. However, achieving high throughput in producing cell-laden microbeads is still a challenge. Our research shows a reliable, single-run approach for the high-throughput generation of hydrogel cell microcarriers (the "cores" of,200 µm) using photo-crosslinking 'on the fly' followed by re-encapsulation in another hydrogel layer to form the "shell" (400-600 µm) for multi-type microtissue engineering. Methods: Firstly, the system included a bead-generation chip with multiple parallel channels. The hydrogel containing a particular cell line was used to form water-in-oil emulsions with a combination of fluorinated oil and surfactant in an external circular track (HFE 7500 and PFPE-PEG-PFPE). The generated microbeads subsequently traveled through a crosslinking chip in front of a LED UV lamp to form crosslinked "cores". Post-washing, the "cores" were remixed with a hydrogel containing the other cell type to create the "shell" in a broadened channel T-junction chip. Results: The system facilitated high throughput and frequency of monodisperse microbeads. The microstructures produced by re-encapsulation showed significant cellular viability (~90 %) with efficient cell compartmentalization. The created platform enabled the culture of the different cell lines to form their niche and maintain crosstalk through the porous hydrogel. The cell-laden droplets are used for micro-tissue engineering to explore various applications. We generated a tumor-stromal model with breast cancer cells forming the tumoral "core" while the "shell" comprised fibroblasts to mimic the tumor microenvironment. Conclusions: In the context of oncology, the 3D cancer-stromal model can help explore the cancer architecture and facilitate the study of the interaction between cancer cells and other components of the tumoral environment. Attempts are currently being made to extend the application of these "core"-"shell" microbeads to generate organoids (artificial pancreatic islets).
ISSN:0391-3988
1724-6040