Cryopreservation of Human iPS Cell Aggregates in a DMSO-Free Solution—An Optimization and Comparative Study

Rui Li, Kathlyn Hornberger, James R. Dutton, Allison Hubel

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Human induced pluripotent stem cells (hiPSCs) are an important cell source for regenerative medicine products. Effective methods of preservation are critical to their clinical and commercial applications. The use of a dimethyl sulfoxide (DMSO)-free solution containing all non-toxic molecules offers an effective alternative to the conventional DMSO and alleviates pain points associated with the use of DMSO in the cryopreservation of hiPSCs. Both hiPSCs and cells differentiated from them are commonly multicellular systems, which are more sensitive to stresses of freezing and thawing than single cells. In this investigation, low-temperature Raman spectroscopy visualized freezing behaviors of hiPSC aggregates in different solutions. These aggregates exhibited sensitivity to undercooling in DMSO-containing solutions. We demonstrated the ability to replace DMSO with non-toxic molecules, improve post-thaw cell survival, and reduce sensitivity to undercooling. An accelerated optimization process capitalized on the positive synergy among multiple DMSO-free molecules, which acted in concert to influence ice formation and protect cells during freezing and thawing. A differential evolution algorithm was used to optimize the multi-variable, DMSO-free preservation protocol in 8 experiments. hiPSC aggregates frozen in the optimized solution did not exhibit the same sensitivity to undercooling as those frozen in non-optimized solutions or DMSO, indicating superior adaptability of the optimized solution to different freezing modalities and unplanned deviations. This investigation shows the importance of optimization, explains the mechanisms and advantages of a DMSO-free solution, and enables not only improved cryopreservation of hiPSCs but potentially other cell types for translational regenerative medicine.

Original languageEnglish (US)
Article number1
JournalFrontiers in Bioengineering and Biotechnology
Volume8
DOIs
StatePublished - Jan 22 2020

Bibliographical note

Funding Information:
This work was supported by the National Institute of Health (R01EB023880 and R25HL128372).

Funding Information:
The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program. The authors thank their colleagues at Ogle laboratory in the Department of Biomedical Engineering, University of Minnesota, for providing the flow cytometer.

Funding Information:
The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program. The authors thank their colleagues at Ogle laboratory in the Department of Biomedical Engineering, University of Minnesota, for providing the flow cytometer. Funding. This work was supported by the National Institute of Health (R01EB023880 and R25HL128372).

Keywords

  • Raman spectroscopy
  • algorithms
  • biomedical engineering
  • cryopreservation
  • human induced pluripotent stem cells

PubMed: MeSH publication types

  • Journal Article

Fingerprint Dive into the research topics of 'Cryopreservation of Human iPS Cell Aggregates in a DMSO-Free Solution—An Optimization and Comparative Study'. Together they form a unique fingerprint.

Cite this