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

Oscar Fabian García-Aponte1, Marta Serra2, Simon Kahlenberg1

  • 1Institute of Cell and Tissue Culture Technology, Department of Biotechnology, BOKU University, Muthgasse 18, Vienna, 1190, Austria.

Advanced Healthcare Materials
|June 8, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a millifluidic method for encapsulating mesenchymal stem cells (MSCs) in microgels, improving their expansion in bioreactors. The process enhances cell viability and expansion by controlling microgel properties and monitoring biological variability.

Keywords:
3D cultureCryo‐SEMencapsulationfluidicshydrogelsmesenchymal stem cells

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Area of Science:

  • Biomaterials Engineering
  • Cell Biology
  • Bioreactor Technology

Background:

  • Mesenchymal stem cells (MSCs) show therapeutic promise but are limited by inefficient, nonphysiological culture methods hindering clinical use.
  • Current cell encapsulation techniques are poorly understood, often neglect biological variability, and inadequately explore dynamic culture conditions.

Purpose of the Study:

  • To develop and standardize a high-throughput millifluidic process for encapsulating MSCs in hydrogel microgels.
  • To investigate the impact of biological variability and crosslinking on MSC expansion under dynamic bioreactor conditions.
  • To assess the potential of this platform for physiologically relevant MSC expansion in large-scale applications.

Main Methods:

  • A high-throughput millifluidic platform was utilized for encapsulating MSCs in gelatin methacryloyl (GelMA) microgels.
  • Cryogenic scanning electron microscopy (Cryo-SEM) was employed to analyze microgel architecture, including the effect of carboxymethyl cellulose.
  • Cell proliferation, microgel shrinkage, glucose uptake, and metabolic activity were monitored to assess MSC expansion and viability under dynamic culture.

Main Results:

  • The millifluidic process yielded highly viable MSC networks within GelMA microgels.
  • Increased crosslinking enhanced microgel resistance to shrinkage, prevented surface cell proliferation, and boosted overall MSC expansion.
  • Interdependencies between cell proliferation, microgel shrinkage, glucose uptake, and cell metabolism were observed, influenced by donor variability.

Conclusions:

  • The developed millifluidic encapsulation platform enables robust and physiologically relevant expansion of MSCs in microgels.
  • Microgel crosslinking is a critical factor in controlling microgel stability and enhancing MSC expansion, even with donor variability.
  • The platform's ability to reliably track metabolic activity alongside cell expansion makes it promising for scalable bioreactor applications.