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Engineering a Microfluidic Platform to Cryopreserve Stem Cells: A DMSO-Free Sustainable Approach.
Saman Modaresi1, Settimio Pacelli2, Aishik Chakraborty3,4
1Department of Chemical and Petroleum Engineering, Bioengineering Graduate Program, School of Engineering, The University of Kansas, Lawrence, KS, 66045, USA.
Advanced Healthcare Materials
|August 17, 2024
Summary
This study introduces a microfluidic chip for DMSO-free cryopreservation of human adipose-derived stem cells (hASCs). The chip enables intracellular delivery of trehalose, a safe alternative, preserving cell function and viability.
Area of Science:
- Biotechnology
- Stem Cell Biology
- Materials Science
Background:
- Traditional cryopreservation of human adipose-derived stem cells (hASCs) relies on dimethyl sulfoxide (DMSO), a cryoprotectant that can induce cytotoxicity, affecting cell proliferation and transplantation outcomes.
- Environmental concerns are associated with the large-scale production and potential contamination of DMSO.
- Trehalose presents a non-cytotoxic, sustainable alternative, but its low cell permeability limits its application in cryopreservation.
Purpose of the Study:
- To develop and validate a microfluidic chip for efficient intracellular delivery of trehalose into hASCs.
- To establish a DMSO-free cryopreservation method for hASCs using trehalose delivered via mechanoporation.
- To assess the cellular compatibility, functionality, and transcriptomic impact of the microfluidic delivery system.
Main Methods:
- Development of a microfluidic chip utilizing mechanoporation to create transient pores in hASC membranes.
- Intracellular delivery of trehalose into hASCs through the microfluidic chip.
- Quantification and optimization of intracellular trehalose delivery based on cellular viability and function.
- Whole-transcriptome sequencing to evaluate gene expression changes in hASCs after microfluidic processing.
Main Results:
- The microfluidic chip successfully enabled intracellular delivery of trehalose in hASCs via mechanoporation.
- Optimized trehalose delivery demonstrated cellular compatibility and maintained hASC functionality.
- Whole-transcriptome sequencing revealed minimal changes (<1% of genes with >2-fold change) in gene expression, indicating low cellular impact.
Conclusions:
- The developed microfluidic chip provides a feasible and effective platform for DMSO-free cryopreservation of hASCs.
- Mechanoporation facilitates intracellular trehalose delivery, offering a safer and potentially more sustainable cryopreservation strategy.
- This approach holds promise for improving the quality and safety of stem cell banking and therapeutic applications.
