Effective surface-based cryopreservation of human embryonic stem cells by vitrification

Beier, A.; Schulz, Julia; Dörr, Daniel; Katsen-Globa, Alisa; Sachinidis, Agapios; Hescheler, Jürgen; Zimmermann, Heiko

doi:10.1016/j.cryobiol.2011.06.003

Cited by 29 publications

(22 citation statements)

References 53 publications

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“…Introduction of i) multi-step freezing, ii) ROCK inhibitors in combination with full cell dissociation, and iii) freezing pluripotent SC's in adherent stage as they are prone to anoikis (cell death after cell are detached from extracellular matrix, [Wagh et al, 2011]) have dramatically increased survival and functionality of human pluripotent cells after SF ( [Katkov et al, 2011;Li et al, 2008;MartinIbanez et al, 2009;Mollamohammadi et al, 2009;Stubban et al, 2007;Ware & Baran, 2007], see Chapter by Martin-Ibanez [Martin-Ibanez, 2012] in this Book). It now highly supersedes various vitrification techniques proposed from time to time [Beier et al, 2011;Reubinoff et al, 2001;Zhou et al, 2004] despite what is claimed otherwise by the authors.…”

Section: Slow Freezing: Still the Mainstream Of Cryopreservation But…mentioning

confidence: 99%

Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature?

Katkov¹,

Bolyukh²,

Chernetsov³

et al. 2012

Current Frontiers in Cryobiology

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Section: Slow Freezing: Still the Mainstream Of Cryopreservation But…mentioning

confidence: 99%

Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature?

Katkov¹,

Bolyukh²,

Chernetsov³

et al. 2012

Current Frontiers in Cryobiology

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“…C, D). Rapid thawing was applied for both freezing methods as previously described . Experiments were performed for unfrozen control cells and cryopreserved cells 1 day (d1) and 4 days (d4) after thawing using ICC, FACS analysis, RNA‐Seq, and scanning electron microscopy (SEM) (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Thus far, disadvantages of vitrification are the complex handling and freezing procedure, and the highly limited number of cells that can be vitrified at the same time . A major advance for vitrification of human pluripotent stem cells was made by further developing a surface‐based approach . Adherent vitrification has been successfully shown for hESCs, using a special disposable allowing cultivation, cryopreservation, storage, and post‐thawing cultivation in a single device (Fig.…”

Section: Introductionmentioning

confidence: 99%

Zooming in on Cryopreservation of hiPSCs and Neural Derivatives: A Dual-Center Study Using Adherent Vitrification

Kaindl

Meiser

Majer

et al. 2018

Stem Cells Translational Medicine

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Human induced pluripotent stem cells (hiPSCs) are an important tool for research and regenerative medicine, but their efficient cryopreservation remains a major challenge. The current gold standard is slow‐rate freezing of dissociated colonies in suspension, but low recovery rates limit immediate post‐thawing applicability. We tested whether ultrafast cooling by adherent vitrification improves post‐thawing survival in a selection of hiPSCs and small molecule neural precursor cells (smNPCs) from Parkinson's disease and controls. In a dual‐center study, we compared the results by immunocytochemistry (ICC), fluorescence‐activated cell sorting analysis, and RNA‐sequencing (RNA‐seq). Adherent vitrification was achieved in the so‐called TWIST substrate, a device combining cultivation, vitrification, storage, and post‐thawing cultivation. Adherent vitrification resulted in preserved confluency and significantly higher cell numbers, and viability at day 1 after thawing, while results were not significantly different at day 4 after thawing. RNA‐seq and ICC of hiPSCs revealed no change in gene expression and pluripotency markers, indicating that physical damage of slow‐rate freezing disrupts cellular membranes. Scanning electron microscopy showed preserved colony integrity by adherent vitrification. Experiments using smNPCs demonstrated that adherent vitrification is also applicable to neural derivatives of hiPSCs. Our data suggest that, compared to the state‐of‐the‐art slow‐rate freezing in suspension, adherent vitrification is an improved cryopreservation technique for hiPSCs and derivatives. Stem Cells Translational Medicine 2019;8:247&259

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“…69 Recently, a surface vitrification method has been developed to vitrify the adherent cell colonies. 70 The cells were immobilized on modified Thermanox ® coverslips with feeder cells before the vitrification, and the vitrified colonies showed better vital residual area compared with slow frozen colonies (89% vs. 51%).…”

Section: Methods Of Hpsc Cryopreservationmentioning

confidence: 98%

Bioprocessing of Cryopreservation for Large-Scale Banking of Human Pluripotent Stem Cells

2012

BioResearch Open Access

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Human pluripotent stem cell (hPSC)-derived cell therapy requires production of therapeutic cells in large quantity, which starts from thawing the cryopreserved cells from a working cell bank or a master cell bank. An optimal cryopreservation and thaw process determines the efficiency of hPSC expansion and plays a significant role in the subsequent lineage-specific differentiation. However, cryopreservation in hPSC bioprocessing has been a challenge due to the unique growth requirements of hPSC, the sensitivity to cryoinjury, and the unscalable cryopreservation procedures commonly used in the laboratory. Tremendous progress has been made to identify the regulatory pathways regulating hPSC responses during cryopreservation and the development of small molecule interventions that effectively improves the efficiency of cryopreservation. The adaption of these methods in current good manufacturing practices (cGMP)-compliant cryopreservation processes not only improves cell survival, but also their therapeutic potency. This review summarizes the advances in these areas and discusses the technical requirements in the development of cGMP-compliant hPSC cryopreservation process.

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Effective surface-based cryopreservation of human embryonic stem cells by vitrification

Cited by 29 publications

References 53 publications

Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature?

Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature?

Zooming in on Cryopreservation of hiPSCs and Neural Derivatives: A Dual-Center Study Using Adherent Vitrification

Bioprocessing of Cryopreservation for Large-Scale Banking of Human Pluripotent Stem Cells

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