Novel cryopreservation method for dissociated human embryonic stem cells in the presence of a ROCK inhibitor
Abstract
Background Human embryonic stem cells (hESCs) have potential use in clinical therapy and regenerative medicine. One of the major challenges regarding the application of these cells is the development of an efficient cryopreservation protocol, since current methods, which include slow-freezing-rapid thawing and vitrification of colonies in suspension, present poor viability and high differentiation rates. Dissociated hESC suspensions do not survive cryopreservation because they are susceptible to apoptosis upon cell detachment and dissociation. A selective Rho-associated kinase (ROCK) inhibitor has been reported to increase the survival of dissociated hESCs and their cloning efficiency. Methods and results Here, we describe a novel method for dissociated hESCs cryopreservation in the presence of the ROCK inhibitor Y-27632. The addition of this inhibitor to the freezing and post-thawing medium significantly increased the survival rate and efficiency of colony formation. Moreover, the hESC colonies obtained after the cryopreservation in the presence of the ROCK inhibitor showed a very low rate of differentiation and a reduced time of recovery. After prolonged culture of frozen-thawed dissociated hESCs, the characteristic properties of pluripotent cells were observed, including normal karyotype, morphological features, marker expression (SSEA-4, TRA-1-60, TRA-1-81 and Oct-4) and the potential to differentiate into derivatives of all three germ layers after embryoid bodies formati…
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
RhoA signalling might potentially contribute to cell death, because it can promote apoptosis in low-adhesion environments by increasing cell contractility.
“RhoA signalling might also potentially contribute to cell death, because it can promote apoptosis in low-adhesion environments by increasing cell contractility, explaining why ROCK inhibitors are a standard component of media for stem cells and organoid isolation and maintenance in culture 77-81 .”
From Taxane-induced conformational changes in the microtubule lattice activate GEF-H1-dependent RhoA signalling (2026) · cites this paper as [77]
No citation lists for this paper
No citation lists for this paper yet — checked again daily.
References 32 · Cited by 128
- Authors
- Martin-Ibanez, R., Unger, C., Stromberg, A., Baker, D., Canals, J. M., & Hovatta, O.
- Affiliations
- Karolinska University Hospital (SE)
- Sheffield Children's NHS Foundation Trust (GB)
- Biomedical Research Networking Center on Neurodegenerative Diseases (ES)
- Venue
- Human Reproduction
- Publisher
- Oxford University Press (OUP)
- Published
- 2008-09-01
- Volume
- 23
- Issue
- 12
- Pages
- 2744-2754
- ISSN
- 0268-1161 (ISSN-L)
- 1460-2350
- OpenAlex
- W2119402851
- Cited by
- 128
- References
- 32
- Citations by year
- 2026: 4 · 2025: 4 · 2024: 1 · 2023: 3 · 2022: 5 · 2021: 1 · 2020: 2 · 2019: 5 · 2018: 5 · 2017: 4 · 2016: 7 · 2015: 4 · 2014: 8 · 2013: 6 · 2012: 11
- FWCI
- 7.95
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Human Reproduction (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Reproductive Biology and Fertility
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Keywords
- human embryonic stem cells, cryopreservation, ROCK inhibitor, Y-27632, pluripotency maintenance, embryoid body differentiation
- MeSH terms
- Amides, Cell Differentiation, Cell Line, Cell Survival, Cryoprotective Agents, Humans, Pyridines, Cryopreservation, Apoptosis, Pluripotent Stem Cells, Embryonic Stem Cells, rho-Associated Kinases
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
