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High-Throughput Encapsulation of Stem Cells: Characterizing Dynamic Culture Variability With a Millifluidic Approach

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Oscar Fabian García-Aponte
  • Marta Serra
  • Simon Kahlenberg
  • Guruprakash Subbiahdoss
  • Dominik Egger

External Research Organisations

  • University of Natural Resources and Applied Life Sciences (BOKU)
  • Universidade de Lisboa

Details

Original languageEnglish
JournalAdvanced healthcare materials
Early online date8 Jun 2025
Publication statusE-pub ahead of print - 8 Jun 2025

Abstract

Mesenchymal stem cells (MSCs) hold potential for several applications, but inefficient nonphysiological culturing methods constantly prevent clinical translation. Automated cell encapsulation in small hydrogels (microgels) facilitates physiologically relevant MSC expansion in bioreactors. Unfortunately, encapsulation processes are poorly characterized, biological variability is seldomly considered, and dynamic culturing is marginally explored. Here, a high-throughput millifluidic encapsulation process is introduced and standardized. This platform enables highly viable MSC networks within gelatin methacryloyl microgels. The impact of biological variability and crosslinking variations under strong dynamic culturing conditions is closely monitored through cell proliferation, microgel shrinkage, and metabolic activity. The effect of carboxymethyl cellulose on microgel's architecture is observed with cryogenic scanning electron microscopy. Increased crosslinking controls the formation of an outer layer on the microgels, which improves the microgel's resistance to shrinking, prevents cell proliferation on the material's surface and increases overall MSC expansion. Cell proliferation, microgel shrinkage, glucose uptake, and cell metabolism show interdependencies observable thanks to the high encapsulation output. Cell proliferation and metabolic activity depend strongly on donor-to-donor variability and change during culture. However, metabolic readouts reliably follow cell expansion, which makes this simple and mechanically robust platform promising for large-scale bioreactor applications.

Keywords

    3D culture, Cryo-SEM, encapsulation, fluidics, hydrogels, mesenchymal stem cells

ASJC Scopus subject areas

Cite this

High-Throughput Encapsulation of Stem Cells: Characterizing Dynamic Culture Variability With a Millifluidic Approach. / García-Aponte, Oscar Fabian; Serra, Marta; Kahlenberg, Simon et al.
In: Advanced healthcare materials, 08.06.2025.

Research output: Contribution to journalArticleResearchpeer review

García-Aponte, O. F., Serra, M., Kahlenberg, S., Subbiahdoss, G., Reimhult, E., Egger, D., & Kasper, C. (2025). High-Throughput Encapsulation of Stem Cells: Characterizing Dynamic Culture Variability With a Millifluidic Approach. Advanced healthcare materials. Advance online publication. https://doi.org/10.1002/adhm.202405137
García-Aponte OF, Serra M, Kahlenberg S, Subbiahdoss G, Reimhult E, Egger D et al. High-Throughput Encapsulation of Stem Cells: Characterizing Dynamic Culture Variability With a Millifluidic Approach. Advanced healthcare materials. 2025 Jun 8. Epub 2025 Jun 8. doi: 10.1002/adhm.202405137
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