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Zooming in on Cryopreservation of hiPSCs and Neural Derivatives: A Dual-Center Study Using Adherent Vitrification
Indexado
WoS WOS:000459820300006
Scopus SCOPUS_ID:85057020352
DOI 10.1002/SCTM.18-0121
Año 2019
Tipo artículo de investigación

Citas Totales

Autores Afiliación Chile

Instituciones Chile

% Participación
Internacional

Autores
Afiliación Extranjera

Instituciones
Extranjeras


Abstract



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.

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Disciplinas de Investigación



WOS
Cell & Tissue Engineering
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 Kaindl, Johanna Mujer FAU Erlangen Nurnberg - Alemania
Friedrich-Alexander-Universität Erlangen-Nürnberg - Alemania
2 Meiser, Ina - Fraunhofer Inst Biomed Engn - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
3 Majer, Julia Mujer Fraunhofer Inst Biomed Engn - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
4 Sommer, Annika Mujer FAU Erlangen Nurnberg - Alemania
Friedrich-Alexander-Universität Erlangen-Nürnberg - Alemania
5 Krach, Florian Hombre FAU Erlangen Nurnberg - Alemania
Univ Calif San Diego - Estados Unidos
Friedrich-Alexander-Universität Erlangen-Nürnberg - Alemania
Department of Cellular & Molecular Medicine - Estados Unidos
6 Katsen-Globa, Alisa Mujer Fraunhofer Inst Biomed Engn - Alemania
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
7 Winkler, Juergen - FAU Erlangen Nurnberg - Alemania
Friedrich-Alexander-Universität Erlangen-Nürnberg - Alemania
8 Zimmermann, Heiko Hombre Fraunhofer Inst Biomed Engn - Alemania
Saarland Univ - Alemania
Universidad Católica del Norte - Chile
Fraunhofer Institute for Biomedical Engineering IBMT - Alemania
Universität des Saarlandes - Alemania
9 Neubauer, Julia C. Mujer Fraunhofer Inst Biomed Engn - 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
10 Winner, Beate Mujer FAU Erlangen Nurnberg - Alemania
Friedrich-Alexander-Universität Erlangen-Nürnberg - Alemania

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Financiamiento



Fuente
Deutsche Forschungsgemeinschaft
DFG
German Federal Ministry of Education and Research
Bundesministerium für Bildung und Forschung
Deutsche Forschungsge-meinschaft
Bundesministerium für Bildung und Forschung
Bavarian California Technology Center (BaCaTec)
DRG
Bavarian Ministry of Economic Affairs, Energy and Technology (grant PROMIB)
Interdisciplinary Centre for Clinical Research (University Hospital of Erlangen E25)
Bavarian Ministry of Education and Culture, Science and the Arts
German Federal Ministry of Education and Research BMBF
Bureau of Energy, Ministry of Economic Affairs, Republic of Taiwan
Bavarian Ministry of Education and Culture
Centre for Interdisciplinary Research in Rehabilitation
Bavarian California Technology Center
Bavarian Ministry of Economic Affairs, Energy and Technology
Interdisciplinary Centre for Clinical Research
Interdisziplinäres Zentrum für Klinische Forschung, Universitätsklinikum Würzburg

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Agradecimientos



Agradecimiento
This work has been funded by the German Federal Ministry of Education and Research BMBF (grant 01EK1609A) and by the Bavarian Ministry of Economic Affairs, Energy and Technology (grant PROMIB). J.K. is an associated member of Research Training Grant GRK2162 of the Deutsche Forschungsgemeinschaft. Additional support came from the Bavarian Ministry of Education and Culture, Science and the Arts in the framework of the Bavarian Research Network Induced Pluripotent Stem Cells for ForIPS, the German Federal Ministry of Education and Research (BMBF: 01GQ113, 01GM1520A), the DFG funded research training group GRK2162 (B.W., J.W.) and DRG grant 410/45-1 FUGG, and the Interdisciplinary Centre for Clinical Research (University Hospital of Erlangen E25). F.K. received funding from the Bavarian California Technology Center (BaCaTec).
This work has been funded by the German Federal Ministry of Education and Research BMBF (grant 01EK1609A) and by the Bavarian Ministry of Economic Affairs, Energy and Technology (grant PROMIB). J.K. is an associated member of Research Training Grant GRK2162 of the Deutsche Forschungsge-meinschaft. Additional support came from the Bavarian Ministry of Education and Culture, Science and the Arts in the framework of the Bavarian Research Network Induced Pluripotent Stem Cells for ForIPS, the German Federal Ministry of Education and Research (BMBF: 01GQ113, 01GM1520A), the DFG funded research training group GRK2162 (B.W., J.W.) and DRG grant 410/45-1 FUGG, and the Interdisciplinary Centre for Clinical Research (University Hospital of Erlangen E25). F.K. received funding from the Bavarian California Technology Center (BaCaTec).

Muestra la fuente de financiamiento declarada en la publicación.