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doi:10.22028/D291-41980
Titel: | Droplet-based vitrification of adherent human induced pluripotent stem cells on alginate microcarrier influenced by adhesion time and matrix elasticity |
VerfasserIn: | Meiser, Ina Majer, Julia Katsen-Globa, Alisa Schulz, André Schmidt, Katharina Stracke, Frank Koutsouraki, Eirini Witt, Gesa Keminer, Oliver Pless, Ole Gardner, John Claussen, Carsten Gribbon, Philip Neubauer, Julia C. Zimmermann, Heiko |
Sprache: | Englisch |
Titel: | Cryobiology |
Bandnummer: | 103 |
Seiten: | 57-69 |
Verlag/Plattform: | Elsevier |
Erscheinungsjahr: | 2021 |
Freie Schlagwörter: | Vitrification Microcarrier Human induced pluripotent stem cells Adherent cryopreservation |
DDC-Sachgruppe: | 500 Naturwissenschaften |
Dokumenttyp: | Journalartikel / Zeitschriftenartikel |
Abstract: | The gold standard in cryopreservation is still conventional slow freezing of single cells or small aggregates in suspension, although major cell loss and limitation to non-specialised cell types in stem cell technology are known drawbacks. The requirement for rapidly available therapeutic and diagnostic cell types is increasing constantly. In the case of human induced pluripotent stem cells (hiPSCs) or their derivates, more sophisticated cryopreservation protocols are needed to address this demand. These should allow a preservation in their physiological, adherent state, an efficient re-cultivation and upscaling upon thawing towards high-throughput applications in cell therapies or disease modelling in drug discovery. Here, we present a novel vitrification based method for adherent hiPSCs, designed for automated handling by microfluidic approaches and with ready-to-use potential e.g. in suspension-based bioreactors after thawing. Modifiable alginate microcarriers serve as a growth surface for adherent hiPSCs that were cultured in a suspension-based bioreactor and subsequently cryopreserved via droplet-based vitrification in comparison to conventional slow freezing. Soft (0.35%) versus stiff (0.65%) alginate microcarriers in concert with adhesion time variation have been examined. Findings revealed specific optimal conditions leading to an adhesion time and growth surface (matrix) elasticity depen dent hypothesis on cryo-induced damaging regimes for adherent cell types. Deviations from the found optimum parameters give rise to membrane ruptures assessed via SEM and major cell loss after adherent vitrification. Applying the optimal conditions, droplet-based vitrification was superior to conventional slow freezing. A decreased microcarrier stiffness was found to outperform stiffer material regarding cell recovery, whereas the stemness characteristics of rewarmed hiPSCs were preserved. |
DOI der Erstveröffentlichung: | 10.1016/j.cryobiol.2021.09.010 |
URL der Erstveröffentlichung: | https://doi.org/10.1016/j.cryobiol.2021.09.010 |
Link zu diesem Datensatz: | urn:nbn:de:bsz:291--ds-419800 hdl:20.500.11880/37566 http://dx.doi.org/10.22028/D291-41980 |
ISSN: | 0011-2240 |
Datum des Eintrags: | 30-Apr-2024 |
Bezeichnung des in Beziehung stehenden Objekts: | Supplementary data |
In Beziehung stehendes Objekt: | https://ars.els-cdn.com/content/image/1-s2.0-S0011224021001681-mmc1.docx |
Fakultät: | NT - Naturwissenschaftlich- Technische Fakultät |
Fachrichtung: | NT - Biowissenschaften |
Professur: | NT - Prof. Dr. Heiko Zimmermann |
Sammlung: | SciDok - Der Wissenschaftsserver der Universität des Saarlandes |
Dateien zu diesem Datensatz:
Datei | Beschreibung | Größe | Format | |
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1-s2.0-S0011224021001681-main.pdf | 18,09 MB | Adobe PDF | Öffnen/Anzeigen |
Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht: Lizenz von Creative Commons