Improving Cell Recovery: Freezing and Thawing Optimization of Induced Pluripotent Stem Cells

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dc.contributor.authorUhrig, Markus
dc.contributor.authorEzquer, Fernando
dc.contributor.authorEzquer, Marcelo
dc.date.accessioned2022-11-14T20:47:04Z
dc.date.available2022-11-14T20:47:04Z
dc.date.issued2022
dc.description.abstractAchieving good cell recovery after cryopreservation is an essential process when working with induced pluripotent stem cells (iPSC). Optimized freezing and thawing methods are required for good cell attachment and survival. In this review, we concentrate on these two aspects, freezing and thawing, but also discuss further factors influencing cell recovery such as cell storage and transport. Whenever a problem occurs during the thawing process of iPSC, it is initially not clear what it is caused by, because there are many factors involved that can contribute to insufficient cell recovery. Thawing problems can usually be solved more quickly when a certain order of steps to be taken is followed. Under optimized conditions, iPSC should be ready for further experiments approximately 4–7 days after thawing and seeding. However, if the freezing and thawing protocols are not optimized, this time can increase up to 2–3 weeks, complicating any further experiments. Here, we suggest optimization steps and troubleshooting options for the freezing, thawing, and seeding of iPSC on feeder-free, Matrigel™-coated, cell culture plates whenever iPSC cannot be recovered in sufficient quality. This review applies to two-dimensional (2D) monolayer cell culture and to iPSC, passaged, frozen, and thawed as cell aggregates (clumps). Furthermore, we discuss usually less well-described factors such as the cell growth phase before freezing and the prevention of osmotic shock during thawing.es
dc.description.versionVersión publicadaes
dc.identifier.citationUhrig, M.; Ezquer, F.; Ezquer, M. Improving Cell Recovery: Freezing and Thawing Optimization of Induced Pluripotent Stem Cells. Cells 2022, 11, 799. https://doi.org/10.3390/cells11050799es
dc.identifier.urihttps://doi.org/10.3390/cells11050799es
dc.identifier.urihttp://hdl.handle.net/11447/6664
dc.language.isoenes
dc.subjectInduced pluripotent stem cellses
dc.subjectLogarithmic cell growth phasees
dc.subjectFreezing protocoles
dc.subjectThawing protocoles
dc.subjectPost-thaw cell recoveryes
dc.subjectiPSC storage and transportes
dc.subjectPreventing osmotic shockes
dc.subjectCell aggregateses
dc.subjectCell seeding densityes
dc.titleImproving Cell Recovery: Freezing and Thawing Optimization of Induced Pluripotent Stem Cellses
dc.typeArticlees
dcterms.sourceCellses

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