Cryopreservation by slow cooling of rat neuronal cells
| Autor: | Claudio Tiribelli, Joaquín V. Rodríguez, Hebe Bottai, Daniel A. Graf, Leonardo Juan de Paz, M. Celeste Robert, Silvia Gazzin |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Male Cell Survival Otras Ciencias Biológicas Biology General Biochemistry Genetics and Molecular Biology Cryopreservation NEURONAL CELLS Ciencias Biológicas Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Cryoprotective Agents Cerebellum Animals Neurons Slow cooling Temperature General Medicine CRYOPRESERVATION SLOW COOLING 030104 developmental biology Female General Agricultural and Biological Sciences Humanities CIENCIAS NATURALES Y EXACTAS 030217 neurology & neurosurgery |
| Zdroj: | Cryobiology. 72(3) |
| ISSN: | 1090-2392 |
| Popis: | Although primary neuronal cells are routinely used for neuroscience research, with potential clinical applications such as neuronal transplantation and tissue engineering, a gold standard protocol for preservation has not been yet developed. In the present work, a slow cooling methodology without ice seeding was studied and optimized for cryopreservation of rat cerebellar granular cells. Parameters such as cooling rate, plunge temperature and cryoprotective agent concentration were assessed using a custom built device based on Pye's freezer idea. Cryopreservation outcome was evaluated by post thawing cell viability/viable cell yield and in culture viability over a period of 14 days. The best outcome was achieved when 10% of Me2SO as cryoprotective agent, a cooling rate of 3.1 ± 0.2 °C/min and a plunge temperature of -48.2 ± 1.5 °C were applied. The granular cells cryopreserved under these conditions exhibited a cell viability of 82.7 ± 2.7% and a viable cell yield of 28.6 ± 2.2%. Moreover, cell viability in culture remained above 50%, very similar to not cryopreserved cells (control). Our results also suggest that post-thaw viability (based on membrane integrity assays) not necessarily reflects the quality of the cryopreservation procedure and proper functionality tests must be carried out in order to optimize both post thaw viability/cell yield and in culture performance. Fil: Robert, María Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Rosario. Secretaria de Ciencia y Técnica. Centro Binacional de Investigación en Criobiología Clínica y Aplicada; Argentina. Fondazione Italiana Fegato; Italia Fil: Juan de Paz, Leonardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Rosario. Secretaria de Ciencia y Técnica. Centro Binacional de Investigación en Criobiología Clínica y Aplicada; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Graf, Daniel A.. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Gazzin, Silvia. Fondazione Italiana Fegato; Italia Fil: Tiribelli, Claudio. Fondazione Italiana Fegato; Italia. University of Trieste; Italia Fil: Bottai, Hebe. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Rodriguez, Joaquin Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Rosario. Secretaria de Ciencia y Técnica. Centro Binacional de Investigación en Criobiología Clínica y Aplicada; Argentina |
| Databáze: | OpenAIRE |
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