SSBD:database

Project ID

472-Okuda-RamanFrozenIPSC

Project SID

472

Title

Raman imaging showing the impact of repeated temperature cycling on cryopreserved human iPSC viability

Description

Human induced pluripotent stem cells (hiPSCs) are an attractive cell source for regenerative medicine. For its widespread use as a starting material, a robust storage and distribution system in the frozen state is necessary. For this system, managing transient warming during storage and transport is essential, but how transient warming affects cells and the mechanisms involved are not yet fully understood. This study examined the influence of temperature cyclings (from -80 deg to -150 deg) on cryopreserved hiPSCs using a custom-made cryo Raman microscope to assess viability. Raman spectroscopy indicated the disappearance of mitochondrial cytochrome signals after thawing. A reduction in the mitochondrial membrane potential was detected using flow cytometry. The performance indices indicated a decrease in attachment effciency with an increase in the number of temperature cycles. This decrease was observed in the temperature cycle range above the glass transition temperature of the cryoprotectant. Raman observations captured an increase in the signal intensity of intracellular dimethyl sulfoxide (DMSO) during temperature cycles. Based on these results, the authors proposed a schematic illustration for cellular responses to temperature fluctuations, suggesting that temperature fluctuations above the glass- transition temperature trigger the movement of DMSO, leading to cytochrome c oxidation, mitochondrial damage, and caspase-mediated cell death. This enhances our understanding of the key events during cryopreservation and informs the development of quality control strategies for hiPSC storage and transport.

Submission Date

2025-09-22

Opened Date

-

Release Date

2026-11-02

Update Date

-

Kind

License

Project URL

-

Project DOI

-

Metadata Version

-

Template Version

3.03

Funding Information

This research was supported by AMED under Grant Number JP20be0704001.

Total

10 files / 2.4 MB

Image datasets

zipped 5 files / 432.0 KB, raw image 5 files / 2.0 MB, total 10 files / 2.4 MB

Quantitative datasets

zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes

Organism

Strain

-

Cell

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Cell line

MeSH

Anatomical Entity

-

UBERON

-

Biological Process

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Cellular Component

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Molecular Function

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Medical Subject Headings

Method involved in biological imaging