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| 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
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.
| 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 |
| 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 |
| 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). |