Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging
Abstract Background Bone marrow-derived mesenchymal stromal cell (MSC) therapy is a promising treatment for several degenerative ocular diseases; however, no reproducible method of monitoring these cells into the eye has been established. The aim of this study was to describe successful bioluminesce...
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BMC
2018-06-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | http://link.springer.com/article/10.1186/s13287-018-0909-y |
| _version_ | 1818294700368461824 |
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| author | Carolina Assis P. Vilela Lucas Eduardo B. Souza Rubens C. Siqueira Rodrigo T. Calado Dimas T. Covas Jayter S. Paula |
| author_facet | Carolina Assis P. Vilela Lucas Eduardo B. Souza Rubens C. Siqueira Rodrigo T. Calado Dimas T. Covas Jayter S. Paula |
| author_sort | Carolina Assis P. Vilela |
| collection | DOAJ |
| description | Abstract Background Bone marrow-derived mesenchymal stromal cell (MSC) therapy is a promising treatment for several degenerative ocular diseases; however, no reproducible method of monitoring these cells into the eye has been established. The aim of this study was to describe successful bioluminescence imaging (BLI) to detect viable luciferase-expressing MSC in the eye. Methods Human donor MSC in culture were transduced with 50 μl luciferase lentiviral vector (three viral particles/cell) prior to intraocular injection. Twenty-one right eyes of 21 rabbits were evaluated through BLI after receiving 1 × 106 luciferase-expressing MSC intravitreally. Contralateral eyes were injected with vehicle (phosphate-buffered saline (PBS)) and were used as controls. At seven different time points (1 h to 60 days), d-luciferin (40 mg/ml, 300 μl PBS) was injected in subsets of six enucleated eyes for evaluation of radiance decay through BLI analysis. CD90 and CD73 immunofluorescence was studied in selected eyes. Results Eyes injected with MSC showed high BLI radiance immediately after d-luciferin injection and progressive decay until 60 days. Mean BLI radiance measures from eyes with luciferase-expressing MSC were significantly higher than controls from 8 h to 30 days. At the thirtieth day, positive CD90- and CD73-expressing cells were observed only in the vitreous cavity of eyes injected with MSC. Conclusions Viable MSC were identified in the vitreous cavity 1 month after a single injection. Our results confirmed BLI as a useful and reliable method to detect MSC injected into the eye globe. |
| first_indexed | 2024-12-13T03:35:55Z |
| format | Article |
| id | doaj.art-c03df426ae9e4003a5118ca150da97ac |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-13T03:35:55Z |
| publishDate | 2018-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-c03df426ae9e4003a5118ca150da97ac2022-12-22T00:01:03ZengBMCStem Cell Research & Therapy1757-65122018-06-01911710.1186/s13287-018-0909-yEx vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imagingCarolina Assis P. Vilela0Lucas Eduardo B. Souza1Rubens C. Siqueira2Rodrigo T. Calado3Dimas T. Covas4Jayter S. Paula5Department of Ophthalmology, Otorhinolaryngology and Head and Neck Surgery, Ribeirão Preto Medical School, University of São PauloHemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo Ribeirão PretoDepartment of Ophthalmology, Otorhinolaryngology and Head and Neck Surgery, Ribeirão Preto Medical School, University of São PauloHemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo Ribeirão PretoHemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo Ribeirão PretoDepartment of Ophthalmology, Otorhinolaryngology and Head and Neck Surgery, Ribeirão Preto Medical School, University of São PauloAbstract Background Bone marrow-derived mesenchymal stromal cell (MSC) therapy is a promising treatment for several degenerative ocular diseases; however, no reproducible method of monitoring these cells into the eye has been established. The aim of this study was to describe successful bioluminescence imaging (BLI) to detect viable luciferase-expressing MSC in the eye. Methods Human donor MSC in culture were transduced with 50 μl luciferase lentiviral vector (three viral particles/cell) prior to intraocular injection. Twenty-one right eyes of 21 rabbits were evaluated through BLI after receiving 1 × 106 luciferase-expressing MSC intravitreally. Contralateral eyes were injected with vehicle (phosphate-buffered saline (PBS)) and were used as controls. At seven different time points (1 h to 60 days), d-luciferin (40 mg/ml, 300 μl PBS) was injected in subsets of six enucleated eyes for evaluation of radiance decay through BLI analysis. CD90 and CD73 immunofluorescence was studied in selected eyes. Results Eyes injected with MSC showed high BLI radiance immediately after d-luciferin injection and progressive decay until 60 days. Mean BLI radiance measures from eyes with luciferase-expressing MSC were significantly higher than controls from 8 h to 30 days. At the thirtieth day, positive CD90- and CD73-expressing cells were observed only in the vitreous cavity of eyes injected with MSC. Conclusions Viable MSC were identified in the vitreous cavity 1 month after a single injection. Our results confirmed BLI as a useful and reliable method to detect MSC injected into the eye globe.http://link.springer.com/article/10.1186/s13287-018-0909-yBioluminescenceCell transplantationCell viability assessmentIntravitreal injectionLuciferaseMesenchymal stromal cell |
| spellingShingle | Carolina Assis P. Vilela Lucas Eduardo B. Souza Rubens C. Siqueira Rodrigo T. Calado Dimas T. Covas Jayter S. Paula Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging Stem Cell Research & Therapy Bioluminescence Cell transplantation Cell viability assessment Intravitreal injection Luciferase Mesenchymal stromal cell |
| title | Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| title_full | Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| title_fullStr | Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| title_full_unstemmed | Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| title_short | Ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| title_sort | ex vivo evaluation of intravitreal mesenchymal stromal cell viability using bioluminescence imaging |
| topic | Bioluminescence Cell transplantation Cell viability assessment Intravitreal injection Luciferase Mesenchymal stromal cell |
| url | http://link.springer.com/article/10.1186/s13287-018-0909-y |
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