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