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Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device
BACKGROUND: A device was devised which aimed to reduce the time and expertise required to perform sonoporation on adherent cell cultures. This prototype device was used to examine the superficial effect of bath temperature on sonoporation efficacy. METHODS: The prototype device consisted of six ultr...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4636885/ https://www.ncbi.nlm.nih.gov/pubmed/26550479 http://dx.doi.org/10.1186/s40349-015-0040-9 |
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author | Kivinen, Jonathan Togtema, Melissa Mulzer, Gregor Choi, Joshua Zehbe, Ingeborg Curiel, Laura Pichardo, Samuel |
author_facet | Kivinen, Jonathan Togtema, Melissa Mulzer, Gregor Choi, Joshua Zehbe, Ingeborg Curiel, Laura Pichardo, Samuel |
author_sort | Kivinen, Jonathan |
collection | PubMed |
description | BACKGROUND: A device was devised which aimed to reduce the time and expertise required to perform sonoporation on adherent cell cultures. This prototype device was used to examine the superficial effect of bath temperature on sonoporation efficacy. METHODS: The prototype device consisted of six ultrasound transducers affixed beneath an Opticell stage. Six transducers with nominal diameters of 20 mm were constructed and the acoustic field of each was characterized using hydrophone scanning. A near field treatment plane was chosen for each transducer to minimize field heterogeneity in the near field. Cervical cancer-derived SiHa cells were exposed to nine different treatments in the presence of plasmid DNA-expressing green fluorescent protein (GFP). Ultrasound treatment with Definity ultrasound contrast agent (US+UCA) present, ultrasound treatment without contrast agent present (US), and a sham ultrasound treatment in the presence of ultrasound contrast agent (CA) were each performed at bath temperatures of 37, 39.5, and 42 °C. Each treatment was performed in biological triplicate. GFP expression and PARP expression following treatment were measured using fluorescent microscopy and digital image processing. Cell detachment was measured using phase contrast microscopy before and after treatment. RESULTS: Mean (± s.d.) transfection rates for the US+UCA treatment were 5.4(±0.92), 5.8(±1.3), and 5.3(±1.1) % at 37, 39.5, and 42 °C, respectively. GFP expression and cell detachment were both significantly affected by the presence of ultrasound contrast agent (p < 0.001, p < 0.001). Neither GFP expression, PARP expression, or detachment differed significantly between bath temperatures. CONCLUSIONS: Bath temperature did not impact the efficacy of sonoporation treatment on SiHa cells in vitro. The prototype device was found to be suitable for performing sonoporation on adherent cell cultures and will reduce the time and expertise required for conducting sonoporation experiments on adherent cell cultures in the future. |
format | Online Article Text |
id | pubmed-4636885 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-46368852015-11-08 Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device Kivinen, Jonathan Togtema, Melissa Mulzer, Gregor Choi, Joshua Zehbe, Ingeborg Curiel, Laura Pichardo, Samuel J Ther Ultrasound Research BACKGROUND: A device was devised which aimed to reduce the time and expertise required to perform sonoporation on adherent cell cultures. This prototype device was used to examine the superficial effect of bath temperature on sonoporation efficacy. METHODS: The prototype device consisted of six ultrasound transducers affixed beneath an Opticell stage. Six transducers with nominal diameters of 20 mm were constructed and the acoustic field of each was characterized using hydrophone scanning. A near field treatment plane was chosen for each transducer to minimize field heterogeneity in the near field. Cervical cancer-derived SiHa cells were exposed to nine different treatments in the presence of plasmid DNA-expressing green fluorescent protein (GFP). Ultrasound treatment with Definity ultrasound contrast agent (US+UCA) present, ultrasound treatment without contrast agent present (US), and a sham ultrasound treatment in the presence of ultrasound contrast agent (CA) were each performed at bath temperatures of 37, 39.5, and 42 °C. Each treatment was performed in biological triplicate. GFP expression and PARP expression following treatment were measured using fluorescent microscopy and digital image processing. Cell detachment was measured using phase contrast microscopy before and after treatment. RESULTS: Mean (± s.d.) transfection rates for the US+UCA treatment were 5.4(±0.92), 5.8(±1.3), and 5.3(±1.1) % at 37, 39.5, and 42 °C, respectively. GFP expression and cell detachment were both significantly affected by the presence of ultrasound contrast agent (p < 0.001, p < 0.001). Neither GFP expression, PARP expression, or detachment differed significantly between bath temperatures. CONCLUSIONS: Bath temperature did not impact the efficacy of sonoporation treatment on SiHa cells in vitro. The prototype device was found to be suitable for performing sonoporation on adherent cell cultures and will reduce the time and expertise required for conducting sonoporation experiments on adherent cell cultures in the future. BioMed Central 2015-11-06 /pmc/articles/PMC4636885/ /pubmed/26550479 http://dx.doi.org/10.1186/s40349-015-0040-9 Text en © Kivinen et al. 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Kivinen, Jonathan Togtema, Melissa Mulzer, Gregor Choi, Joshua Zehbe, Ingeborg Curiel, Laura Pichardo, Samuel Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title | Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title_full | Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title_fullStr | Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title_full_unstemmed | Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title_short | Sonoporation efficacy on SiHa cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
title_sort | sonoporation efficacy on siha cells in vitro at raised bath temperatures—experimental validation of a prototype sonoporation device |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4636885/ https://www.ncbi.nlm.nih.gov/pubmed/26550479 http://dx.doi.org/10.1186/s40349-015-0040-9 |
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