A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment
Low oxygen tensions experienced in various pathological and physiological conditions are a major stimulus for angiogenesis. Hypoxic conditions play a critical role in regulating cellular behaviour including migration, proliferation and differentiation. This study introduces the use of a microfluidic...
| Principais autores: | , , , , , |
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| Formato: | Artigo |
| Idioma: | en_US |
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Royal Society of Chemistry
2014
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| Acesso em linha: | http://hdl.handle.net/1721.1/89411 https://orcid.org/0000-0002-7232-304X |
| _version_ | 1826189777086447616 |
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| author | Funamoto, Kenichi Zervantonakis, Ioannis K. Liu, Yuchun Ochs, Christopher J. Kim, Choong Kamm, Roger Dale |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Funamoto, Kenichi Zervantonakis, Ioannis K. Liu, Yuchun Ochs, Christopher J. Kim, Choong Kamm, Roger Dale |
| author_sort | Funamoto, Kenichi |
| collection | MIT |
| description | Low oxygen tensions experienced in various pathological and physiological conditions are a major stimulus for angiogenesis. Hypoxic conditions play a critical role in regulating cellular behaviour including migration, proliferation and differentiation. This study introduces the use of a microfluidic device that allows for the control of oxygen tension for the study of different three-dimensional (3D) cell cultures for various applications. The device has a central 3D gel region acting as an external cellular matrix, flanked by media channels. On each side, there is a peripheral gas channel through which suitable gas mixtures are supplied to establish a uniform oxygen tension or gradient within the device. The effects of various parameters, such as gas and media flow rates, device thickness, and diffusion coefficients of oxygen were examined using numerical simulations to determine the characteristics of the microfluidic device. A polycarbonate (PC) film with a low oxygen diffusion coefficient was embedded in the device in proximity above the channels to prevent oxygen diffusion from the incubator environment into the polydimethylsiloxane (PDMS) device. The oxygen tension in the device was then validated experimentally using a ruthenium-coated (Ru-coated) oxygen-sensing glass cover slip which confirmed the establishment of low uniform oxygen tensions (<3%) or an oxygen gradient across the gel region. To demonstrate the utility of the microfluidic device for cellular experiments under hypoxic conditions, migratory studies of MDA-MB-231 human breast cancer cells were performed. The microfluidic device allowed for imaging cellular migration with high-resolution, exhibiting an enhanced migration in hypoxia in comparison to normoxia. This microfluidic device presents itself as a promising platform for the investigation of cellular behaviour in a 3D gel scaffold under varying hypoxic conditions. |
| first_indexed | 2024-09-23T08:21:28Z |
| format | Article |
| id | mit-1721.1/89411 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:21:28Z |
| publishDate | 2014 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | mit-1721.1/894112022-09-23T12:27:15Z A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment Funamoto, Kenichi Zervantonakis, Ioannis K. Liu, Yuchun Ochs, Christopher J. Kim, Choong Kamm, Roger Dale Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Mechanical Engineering Funamoto, Kenichi Zervantonakis, Ioannis K. Kamm, Roger Dale Low oxygen tensions experienced in various pathological and physiological conditions are a major stimulus for angiogenesis. Hypoxic conditions play a critical role in regulating cellular behaviour including migration, proliferation and differentiation. This study introduces the use of a microfluidic device that allows for the control of oxygen tension for the study of different three-dimensional (3D) cell cultures for various applications. The device has a central 3D gel region acting as an external cellular matrix, flanked by media channels. On each side, there is a peripheral gas channel through which suitable gas mixtures are supplied to establish a uniform oxygen tension or gradient within the device. The effects of various parameters, such as gas and media flow rates, device thickness, and diffusion coefficients of oxygen were examined using numerical simulations to determine the characteristics of the microfluidic device. A polycarbonate (PC) film with a low oxygen diffusion coefficient was embedded in the device in proximity above the channels to prevent oxygen diffusion from the incubator environment into the polydimethylsiloxane (PDMS) device. The oxygen tension in the device was then validated experimentally using a ruthenium-coated (Ru-coated) oxygen-sensing glass cover slip which confirmed the establishment of low uniform oxygen tensions (<3%) or an oxygen gradient across the gel region. To demonstrate the utility of the microfluidic device for cellular experiments under hypoxic conditions, migratory studies of MDA-MB-231 human breast cancer cells were performed. The microfluidic device allowed for imaging cellular migration with high-resolution, exhibiting an enhanced migration in hypoxia in comparison to normoxia. This microfluidic device presents itself as a promising platform for the investigation of cellular behaviour in a 3D gel scaffold under varying hypoxic conditions. 2014-09-10T15:38:33Z 2014-09-10T15:38:33Z 2012-09 2012-03 Article http://purl.org/eprint/type/JournalArticle 1473-0197 1473-0189 http://hdl.handle.net/1721.1/89411 Funamoto, Kenichi, Ioannis K. Zervantonakis, Yuchun Liu, Christopher J. Ochs, Choong Kim, and Roger D. Kamm. “A Novel Microfluidic Platform for High-Resolution Imaging of a Three-Dimensional Cell Culture Under a Controlled Hypoxic Environment.” Lab Chip 12, no. 22 (2012): 4855. https://orcid.org/0000-0002-7232-304X en_US http://dx.doi.org/10.1039/c2lc40306d Lab on a Chip Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Royal Society of Chemistry PMC |
| spellingShingle | Funamoto, Kenichi Zervantonakis, Ioannis K. Liu, Yuchun Ochs, Christopher J. Kim, Choong Kamm, Roger Dale A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title | A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title_full | A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title_fullStr | A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title_full_unstemmed | A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title_short | A novel microfluidic platform for high-resolution imaging of a three-dimensional cell culture under a controlled hypoxic environment |
| title_sort | novel microfluidic platform for high resolution imaging of a three dimensional cell culture under a controlled hypoxic environment |
| url | http://hdl.handle.net/1721.1/89411 https://orcid.org/0000-0002-7232-304X |
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