A high-throughput microfluidic device to study neurite response to growth factor gradients
Studying neurite guidance by diffusible or substrate bound gradients is challenging with current techniques. In this study, we present the design, fabrication and utility of a microfluidic device to study neurite guidance under chemogradients. Experimental and computational studies demonstrated the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Royal Society of Chemistry
2012
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| Online Access: | http://hdl.handle.net/1721.1/69153 https://orcid.org/0000-0003-3214-4554 https://orcid.org/0000-0002-7232-304X |
| _version_ | 1826195482963083264 |
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| author | Kothapalli, Chandrasekhar R. de Valence, Sarra Chung, Seok Zervantonakis, Ioannis Gertler, Frank Kamm, Roger Dale Van Veen, John |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Kothapalli, Chandrasekhar R. de Valence, Sarra Chung, Seok Zervantonakis, Ioannis Gertler, Frank Kamm, Roger Dale Van Veen, John |
| author_sort | Kothapalli, Chandrasekhar R. |
| collection | MIT |
| description | Studying neurite guidance by diffusible or substrate bound gradients is challenging with current techniques. In this study, we present the design, fabrication and utility of a microfluidic device to study neurite guidance under chemogradients. Experimental and computational studies demonstrated the establishment of a steep gradient of guidance cue within 30 min and stable for up to 48 h. The gradient was found to be insensitive to external perturbations such as media change and movement of device. The effects of netrin-1 (0.1–10 µg mL−1) and brain pulp (0.1 µL mL−1) were evaluated for their chemoattractive potential on neurite turning, while slit-2 (62.5 or 250 ng mL−1) was studied for its chemorepellant properties. Hippocampal or dorsal root ganglion (DRG) neurons were seeded into a micro-channel and packed onto the surface of a 3D collagen gel. Neurites grew into the matrix in three dimensions, and a gradient of guidance cue was created orthogonal to the direction of neurite growth to impact guidance. The average turning angle of each neurite was measured and averaged across multiple devices cultured under similar conditions to quantify the effect of guidance cue gradient. Significant positive turning towards gradient was measured in the presence of brain pulp and netrin-1 (1 µg mL−1), relative to control cultures which received no external guidance cue (p < 0.001). Netrin-1 released from transfected fibroblasts had the most positive turning effect of all the chemoattractive cues tested (p < 0.001). Slit-2 exhibited strong chemorepellant characteristics on both hippocampal and DRG neurite guidance at 250 ng mL−1 concentration. Slit-2 also showed similar behavior on DRG neuron invasion into 3D collagen gel (p < 0.01 relative to control cultures). Taken together, the results suggest the utility of this microfluidic device to generate stable chemogradients for studying neurobiology, cell migration and proliferation, matrix remodeling and co-cultures with other cell lines, with potential applications in cancer biology, tissue engineering and regenerative medicine. |
| first_indexed | 2024-09-23T10:13:23Z |
| format | Article |
| id | mit-1721.1/69153 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:13:23Z |
| publishDate | 2012 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | mit-1721.1/691532022-09-26T16:33:33Z A high-throughput microfluidic device to study neurite response to growth factor gradients A high-throughput microfluidic assay to study neurite response to growth factor gradients Kothapalli, Chandrasekhar R. de Valence, Sarra Chung, Seok Zervantonakis, Ioannis Gertler, Frank Kamm, Roger Dale Van Veen, John Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Mechanical Engineering Kamm, Roger Dale Kamm, Roger Dale Kothapalli, Chandrasekhar R. Van Veen, Ed de Valence, Sarra Zervantonakis, Ioannis Gertler, Frank Studying neurite guidance by diffusible or substrate bound gradients is challenging with current techniques. In this study, we present the design, fabrication and utility of a microfluidic device to study neurite guidance under chemogradients. Experimental and computational studies demonstrated the establishment of a steep gradient of guidance cue within 30 min and stable for up to 48 h. The gradient was found to be insensitive to external perturbations such as media change and movement of device. The effects of netrin-1 (0.1–10 µg mL−1) and brain pulp (0.1 µL mL−1) were evaluated for their chemoattractive potential on neurite turning, while slit-2 (62.5 or 250 ng mL−1) was studied for its chemorepellant properties. Hippocampal or dorsal root ganglion (DRG) neurons were seeded into a micro-channel and packed onto the surface of a 3D collagen gel. Neurites grew into the matrix in three dimensions, and a gradient of guidance cue was created orthogonal to the direction of neurite growth to impact guidance. The average turning angle of each neurite was measured and averaged across multiple devices cultured under similar conditions to quantify the effect of guidance cue gradient. Significant positive turning towards gradient was measured in the presence of brain pulp and netrin-1 (1 µg mL−1), relative to control cultures which received no external guidance cue (p < 0.001). Netrin-1 released from transfected fibroblasts had the most positive turning effect of all the chemoattractive cues tested (p < 0.001). Slit-2 exhibited strong chemorepellant characteristics on both hippocampal and DRG neurite guidance at 250 ng mL−1 concentration. Slit-2 also showed similar behavior on DRG neuron invasion into 3D collagen gel (p < 0.01 relative to control cultures). Taken together, the results suggest the utility of this microfluidic device to generate stable chemogradients for studying neurobiology, cell migration and proliferation, matrix remodeling and co-cultures with other cell lines, with potential applications in cancer biology, tissue engineering and regenerative medicine. Seoul R&BD Support Center (program PA090930) 2012-02-22T16:18:05Z 2012-02-22T16:18:05Z 2010-11 2010-07 Article http://purl.org/eprint/type/JournalArticle 1473-0197 1473-0189 http://hdl.handle.net/1721.1/69153 Kothapalli, Chandrasekhar R. et al. “A High-throughput Microfluidic Assay to Study Neurite Response to Growth Factor Gradients.” Lab on a Chip 11.3 (2011): 497. https://orcid.org/0000-0003-3214-4554 https://orcid.org/0000-0002-7232-304X en_US http://dx.doi.org/10.1039/c0lc00240b Lab on a Chip Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Royal Society of Chemistry Prof. Kamm via Angie Locknar |
| spellingShingle | Kothapalli, Chandrasekhar R. de Valence, Sarra Chung, Seok Zervantonakis, Ioannis Gertler, Frank Kamm, Roger Dale Van Veen, John A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title | A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title_full | A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title_fullStr | A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title_full_unstemmed | A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title_short | A high-throughput microfluidic device to study neurite response to growth factor gradients |
| title_sort | high throughput microfluidic device to study neurite response to growth factor gradients |
| url | http://hdl.handle.net/1721.1/69153 https://orcid.org/0000-0003-3214-4554 https://orcid.org/0000-0002-7232-304X |
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