Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it>
<p>Abstract</p> <p>Live animal imaging is becoming an increasingly common technique for accurate and quantitative assessment of tumor burden over time. Bioluminescence imaging systems rely on a bioluminescent signal from tumor cells, typically generated from expression of the firef...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-11-01
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| Series: | Molecular Cancer |
| Online Access: | http://www.molecular-cancer.com/content/9/1/299 |
| _version_ | 1818826783794921472 |
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| author | Rasko John EJ Ng Cynthia Bailey Charles G Tiffen Jessamy C Holst Jeff |
| author_facet | Rasko John EJ Ng Cynthia Bailey Charles G Tiffen Jessamy C Holst Jeff |
| author_sort | Rasko John EJ |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Live animal imaging is becoming an increasingly common technique for accurate and quantitative assessment of tumor burden over time. Bioluminescence imaging systems rely on a bioluminescent signal from tumor cells, typically generated from expression of the firefly luciferase gene. However, previous reports have suggested that either a high level of luciferase or the resultant light reaction produced upon addition of D-luciferin substrate can have a negative influence on tumor cell growth. To address this issue, we designed an expression vector that allows simultaneous fluorescence and luminescence imaging. Using fluorescence activated cell sorting (FACS), we generated clonal cell populations from a human breast cancer (MCF-7) and a mouse melanoma (B16-F10) cell line that stably expressed different levels of luciferase. We then compared the growth capabilities of these clones <it>in vitro </it>by MTT proliferation assay and <it>in vivo </it>by bioluminescence imaging of tumor growth in live mice. Surprisingly, we found that neither the amount of luciferase nor biophotonic activity was sufficient to inhibit tumor cell growth, <it>in vitro </it>or <it>in vivo</it>. These results suggest that luciferase toxicity is not a necessary consideration when designing bioluminescence experiments, and therefore our approach can be used to rapidly generate high levels of luciferase expression for sensitive imaging experiments.</p> |
| first_indexed | 2024-12-19T00:33:09Z |
| format | Article |
| id | doaj.art-266c1c12f4c342d89fd490d887c50bc1 |
| institution | Directory Open Access Journal |
| issn | 1476-4598 |
| language | English |
| last_indexed | 2024-12-19T00:33:09Z |
| publishDate | 2010-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Cancer |
| spelling | doaj.art-266c1c12f4c342d89fd490d887c50bc12022-12-21T20:45:00ZengBMCMolecular Cancer1476-45982010-11-019129910.1186/1476-4598-9-299Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it>Rasko John EJNg CynthiaBailey Charles GTiffen Jessamy CHolst Jeff<p>Abstract</p> <p>Live animal imaging is becoming an increasingly common technique for accurate and quantitative assessment of tumor burden over time. Bioluminescence imaging systems rely on a bioluminescent signal from tumor cells, typically generated from expression of the firefly luciferase gene. However, previous reports have suggested that either a high level of luciferase or the resultant light reaction produced upon addition of D-luciferin substrate can have a negative influence on tumor cell growth. To address this issue, we designed an expression vector that allows simultaneous fluorescence and luminescence imaging. Using fluorescence activated cell sorting (FACS), we generated clonal cell populations from a human breast cancer (MCF-7) and a mouse melanoma (B16-F10) cell line that stably expressed different levels of luciferase. We then compared the growth capabilities of these clones <it>in vitro </it>by MTT proliferation assay and <it>in vivo </it>by bioluminescence imaging of tumor growth in live mice. Surprisingly, we found that neither the amount of luciferase nor biophotonic activity was sufficient to inhibit tumor cell growth, <it>in vitro </it>or <it>in vivo</it>. These results suggest that luciferase toxicity is not a necessary consideration when designing bioluminescence experiments, and therefore our approach can be used to rapidly generate high levels of luciferase expression for sensitive imaging experiments.</p>http://www.molecular-cancer.com/content/9/1/299 |
| spellingShingle | Rasko John EJ Ng Cynthia Bailey Charles G Tiffen Jessamy C Holst Jeff Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> Molecular Cancer |
| title | Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> |
| title_full | Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> |
| title_fullStr | Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> |
| title_full_unstemmed | Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> |
| title_short | Luciferase expression and bioluminescence does not affect tumor cell growth <it>in vitro </it>or <it>in vivo</it> |
| title_sort | luciferase expression and bioluminescence does not affect tumor cell growth it in vitro it or it in vivo it |
| url | http://www.molecular-cancer.com/content/9/1/299 |
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