Evidence map›Paper›PMID 40495211›Full record

ArticleStem cell research & therapy2025

DMSO-free cryopreservation of hiPSC-derived cardiomyocytes: low temperature characterization and protocol development.

Akshat S Mallya, Tessa Burrows, Jeanne Hsieh, Troy Louwagie, James R Dutton, Brenda M Ogle, Allison Hubel

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Stem cell research & therapy, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Article
  2. Review
  3. Review
4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

7 authors.

Akshat S MallyaDepartment of Mechanical Engineering, University of Minnesota, Minneapolis, MN, USA.
Tessa BurrowsDepartment of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA.
Jeanne HsiehDepartment of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA.
Troy LouwagieDepartment of Mechanical Engineering, University of Minnesota, Minneapolis, MN, USA.
James R DuttonDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA.
Brenda M OgleDepartment of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA.
Allison HubelDepartment of Mechanical Engineering, University of Minnesota, Minneapolis, MN, USA. hubel001@umn.edu.ORCID http://orcid.org/0000-0001-5856-0500

Funding

Multicomponent solutions for the preservation of cell therapy productsR01HL154734 · NHLBI · UNIVERSITY OF MINNESOTA · PI HUBEL, ALLISON · 2020 to 2023
$1.6M
NHLBI NIH HHS R01 HL154734NIH HHS R01HL154734
6 · The paper itself

Abstract

backgroundHuman-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have attracted significant interest for use in disease modeling, drug discovery and potential therapeutic applications. However, conventional hiPSC-CM cryopreservation protocols largely use dimethyl sulfoxide (DMSO) as the cryoprotectant (CPA), which is linked with a loss of post-thaw recovery and function for various cell types and is not ideal for therapeutic protocols. Additionally, the effect of freezing parameters such as cooling rate and nucleation temperature on post-thaw recovery of hiPSC-CMs has not been explored.

methodshiPSC-CMs were generated by Wnt pathway inhibition, followed by sodium l-lactate purification. Subsequently, biophysical characterization of the cells was performed. A differential evolution (DE) algorithm was utilized to determine the optimal composition of a mixture of a sugar, sugar alcohol and amino acid to replace DMSO as the CPA. The hiPSC-CMs were subjected to controlled-rate freezing at different cooling rates and nucleation temperatures. The optimum freezing parameters were identified by post-thaw recoveries and the partitioning ratio obtained from low temperature Raman spectroscopy studies. The post-thaw osmotic behavior of hiPSC-CMs was studied by measuring diameter of cells resuspended in the isotonic culture medium over time. Immunocytochemistry and calcium transient studies were performed to evaluate post-thaw function.

resultshiPSC-CMs were found to be slightly larger than hiPSCs and exhibited a large osmotically inactive volume. The best-performing DMSO-free solutions enabled post-thaw recoveries over 90%, which was significantly greater than DMSO (69.4 ± 6.4%). A rapid cooling rate of 5 °C/min and a low nucleation temperature of -8 °C was found to be optimal for hiPSC-CMs. hiPSC-CMs displayed anomalous osmotic behavior post-thaw, dropping sharply in volume after resuspension. Post-thaw function was preserved when hiPSC-CMs were frozen with the best-performing DMSO-free CPA or DMSO and the cells displayed similar cardiac markers pre-freeze and post-thaw.

conclusionsIt was shown that a CPA cocktail of naturally-occurring osmolytes could effectively replace DMSO for preserving hiPSC-CMs while preserving morphology and function. Understanding the anomalous osmotic behavior and managing the excessive dehydration of hiPSC-CMs could be crucial to improve post-thaw outcomes. Effective DMSO-free cryopreservation would accelerate the development of drug discovery and therapeutic applications of hiPSC-CMs.

Indexed as

CryopreservationInduced Pluripotent Stem CellsMyocytes, CardiacCell DifferentiationCells, CulturedCryoprotective AgentsDimethyl SulfoxideHumansCryoprotective AgentsDimethyl SulfoxideCardiomyocytesCryopreservationDMSO-freeNADESProtocol developmentRaman spectroscopy

Identifiers

PMID40495211
PMCPMC12150479

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.