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Intracellular loading of cryoprotective agents (CPAs) into target cells is a critical step for cryopreservation. However, biological membranes are usually much less permeable to CPAs than to water, resulting in high osmotic pressures and osmotic damage during the CPA loading and unloading phases of cryopreservation. Here, we show that calcium alginate hydrogel beads several millimeters in diamater containing CPAs can be admixed with a cell suspension to spontaneously release CPAs in a gradual and distributed manner. We demonstrate that beads containing cell media enable the gradual removal of CPA from Jurkat cells equilibrated in a typical cryopreservation solution of 15% glycerol, protecting the cells from hypotonic damage. We show that the dynamics of CPA exchange are accurately described by a numerical model of free diffusion within the gel. This approach may enable semiautomated and closed methods of gradual CPA exchange from large volume cell suspensions.
|Original language||English (US)|
|Number of pages||3|
|State||Published - Dec 2021|
Bibliographical noteFunding Information:
The authors wish to thank Jon Edd and Lindong Weng for helpful discussions and feedback. This work was generously supported by NIH R01DK114506 and NSF EEC1941543 .
© 2021 Elsevier Inc.
- Dimethyl sulfoxide
- Osmotic shock
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, U.S. Gov't, Non-P.H.S.
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- 1 Active
ATP-Bio: NSF Engineering Research Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio)
Bischof, J. C., Toner, M., Roehrig, G. H., Aguilar, G. & Healy, K. E.
National Science Foundation, NSF
9/1/20 → 8/31/25
Project: Research project