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Droplet-based vitrification of adherent human induced pluripotent stem cells on alginate microcarrier influenced by adhesion time and matrix elasticity
Indexado
WoS WOS:000741499100009
Scopus SCOPUS_ID:85116363773
DOI 10.1016/J.CRYOBIOL.2021.09.010
Año 2021
Tipo artículo de investigación

Citas Totales

Autores Afiliación Chile

Instituciones Chile

% Participación
Internacional

Autores
Afiliación Extranjera

Instituciones
Extranjeras


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 vitrificationbased 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 dependent 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.

Revista



Revista ISSN
Cryobiology 0011-2240

Métricas Externas



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Disciplinas de Investigación



WOS
Physiology
Biology
Scopus
Sin Disciplinas
SciELO
Sin Disciplinas

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Publicaciones WoS (Ediciones: ISSHP, ISTP, AHCI, SSCI, SCI), Scopus, SciELO Chile.

Colaboración Institucional



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Autores - Afiliación



Ord. Autor Género Institución - País
1 Meiser, Ina - Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
2 Majer, Julia Mujer Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
3 Katsen-Globa, Alisa Mujer Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
4 Schulz, Andre Hombre Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
5 Schmidt, Katharina Mujer Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
6 Stracke, Frank Hombre Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
7 Koutsouraki, Eirini Mujer Censo Biotechnol Ltd - Reino Unido
Censo Biotechnologies Ltd - Reino Unido
8 Witt, Gesa Mujer ScreeningPort - Alemania
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP - Alemania
9 Keminer, Oliver Hombre ScreeningPort - Alemania
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP - Alemania
10 Pless, Ole - ScreeningPort - Alemania
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP - Alemania
11 Gardner, John Hombre Censo Biotechnol Ltd - Reino Unido
Censo Biotechnologies Ltd - Reino Unido
12 Claussen, Carsten Hombre ScreeningPort - Alemania
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP - Alemania
13 Gribbon, Philip Hombre ScreeningPort - Alemania
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP - Alemania
14 Neubauer, Julia C. Mujer Fraunhofer Inst Biomed Engn IBMT - Alemania
Fraunhofer Project Ctr Stem Cell Proc Engn - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
Fraunhofer Project Centre for Stem Cell Process Engineering - Alemania
15 Zimmermann, Heiko Hombre Fraunhofer Inst Biomed Engn IBMT - Alemania
Censo Biotechnol Ltd - Reino Unido
Universidad Católica del Norte - Chile
Saarland Univ - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
Universität des Saarlandes - Alemania
Censo Biotechnologies Ltd - Reino Unido

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Financiamiento



Fuente
European Commission
Seventh Framework Programme
FP7 Health
Seventh Framework Programme (FP7) of the European Commission (FP7-HEALTH)

Muestra la fuente de financiamiento declarada en la publicación.

Agradecimientos



Agradecimiento
This study was funded under the Seventh Framework Programme (FP7) of the European Commission (FP7-HEALTH), grant agreement: 601865 (DropTech). We thank Dr. Rachel Steeg for careful proofreading.
This study was funded under the Seventh Framework Programme (FP7) of the European Commission ( FP7-HEALTH ), grant agreement: 601865 (DropTech). We thank Dr. Rachel Steeg for careful proofreading.

Muestra la fuente de financiamiento declarada en la publicación.