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. 2023 Oct 21;14:6685. doi: 10.1038/s41467-023-42297-0

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Schematic of triple-jet in-air microfluidic cell encapsulation in microcapsules. b Viability quantification of encapsulated 3T3 fibroblasts with and without 20x dilution upon collection (n = 85 microcapsules). Data is presented as mean values ± SD. Microphotographs of c solid microgels and d hollow microcapsules containing 10⁷ 3T3 fibroblasts/ml after 0 and 7 days culture. e Quantification and microphotographs of 3T3 fibroblast spheroid diameter within microcapsules during 7 days of culture (n = 60, p = 1.37E-29). Data is presented as mean values ± SD. f Confocal fluorescence microphotograph of a microcapsule and its 3T3 spheroid of which the nuclei are stained with DAPI (blue) and its F-actin is stained with phalloidin (green) after 14 days of culture. Microphotographs of spheroids of which their respective microcapsule was instantly dissolved using g PBS or h alginate lyase after 14 days of culture. Microphotographs of i non-merging encapsulated spheroids and j merging de-encapsulated spheroids. k Microphotographs of droplet formation using solutions containing 0, 2*10⁶ and 10⁷ cells/ml. l Quantification of cells per microcapsule for various cell concentrations ranging up to 10⁷ cells/ml (n = 60). Data is presented as mean values ± SD. m Encapsulation efficiency of microcapsule production using 2*10⁶ or 10⁷ cells/ml was quantified (n = 60, p = 1.01E-4). Data is presented as mean values ± SD. n Microphotograph of conventional oil-based microfluidic droplet formation using oil and surfactants, microphotograph of samples obtained with oil-based microfluidics using Oil Red O (ORO) stained oil, and absorbance at 521 nm for residue oil quantification. The chip-based sample was obtained through three thorough washing steps (n = 3). Data is presented as mean values ± SD. Significance was determined based on one-way ANOVA analysis. The significance of p < 0.05 is indicated by *. A.U. Indicates arbitrary units. Scale bar is 200 µm unless stated otherwise.