Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina
AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs) in a mouse model of laser-induced retinal injury. METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluore...
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| Format: | Article |
| Language: | English |
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Press of International Journal of Ophthalmology (IJO PRESS)
2018-04-01
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| Series: | International Journal of Ophthalmology |
| Subjects: | |
| Online Access: | http://www.ijo.cn/en_publish/2018/4/20180407.pdf |
| _version_ | 1819073518878326784 |
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| author | Hui Shi Xin-Rui Wang Ming-Chao Bi Wei Yang Dan Wang Hai-Le Liu Ling-Ling Liang Xiao-Hong Li Qian Hao Zhi-Hua Cui E Song |
| author_facet | Hui Shi Xin-Rui Wang Ming-Chao Bi Wei Yang Dan Wang Hai-Le Liu Ling-Ling Liang Xiao-Hong Li Qian Hao Zhi-Hua Cui E Song |
| author_sort | Hui Shi |
| collection | DOAJ |
| description | AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs) in a mouse model of laser-induced retinal injury.
METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein (DiI-AcLDL), and green fluorescent protein (GFP). The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo.
RESULTS: EPCs labeled with CFSE and DiI-AcLDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and DiI-AcLDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina.
CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and DiI-AcLDL are suitable for short-term EPC-labeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study. |
| first_indexed | 2024-12-21T17:54:54Z |
| format | Article |
| id | doaj.art-42ec4deffb8249a58e6dff82f4b108fe |
| institution | Directory Open Access Journal |
| issn | 2222-3959 2227-4898 |
| language | English |
| last_indexed | 2024-12-21T17:54:54Z |
| publishDate | 2018-04-01 |
| publisher | Press of International Journal of Ophthalmology (IJO PRESS) |
| record_format | Article |
| series | International Journal of Ophthalmology |
| spelling | doaj.art-42ec4deffb8249a58e6dff82f4b108fe2022-12-21T18:55:14ZengPress of International Journal of Ophthalmology (IJO PRESS)International Journal of Ophthalmology2222-39592227-48982018-04-0111458058810.18240/ijo.2018.04.07Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retinaHui Shi0Xin-Rui Wang1Ming-Chao Bi2Wei Yang3Dan Wang4Hai-Le Liu5Ling-Ling Liang6Xiao-Hong Li7Qian Hao8Zhi-Hua Cui9E Song10Department of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaKey Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaDepartment of Ophthalmology, Lixiang Eye Hospital, Soochow University, Suzhou 215021, Jiangsu Province, China; Department of Ophthalmology, First Hospital, Jilin University, Changchun 130021, Jilin Province, ChinaAIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs) in a mouse model of laser-induced retinal injury. METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE), 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein (DiI-AcLDL), and green fluorescent protein (GFP). The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo. RESULTS: EPCs labeled with CFSE and DiI-AcLDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and DiI-AcLDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina. CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and DiI-AcLDL are suitable for short-term EPC-labeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.http://www.ijo.cn/en_publish/2018/4/20180407.pdf588endothelial progenitor cellscell tracking5-(and-6)- carboxyfluorescein diacetate succinimidyl ester11′-dilinoleyl-333′3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoproteingreen fluorescent proteinretinal laser photocoagulation |
| spellingShingle | Hui Shi Xin-Rui Wang Ming-Chao Bi Wei Yang Dan Wang Hai-Le Liu Ling-Ling Liang Xiao-Hong Li Qian Hao Zhi-Hua Cui E Song Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina International Journal of Ophthalmology 588 endothelial progenitor cells cell tracking 5-(and-6)- carboxyfluorescein diacetate succinimidyl ester 1 1′-dilinoleyl-3 3 3′ 3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein green fluorescent protein retinal laser photocoagulation |
| title | Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina |
| title_full | Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina |
| title_fullStr | Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina |
| title_full_unstemmed | Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina |
| title_short | Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina |
| title_sort | comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser injured retina |
| topic | 588 endothelial progenitor cells cell tracking 5-(and-6)- carboxyfluorescein diacetate succinimidyl ester 1 1′-dilinoleyl-3 3 3′ 3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein green fluorescent protein retinal laser photocoagulation |
| url | http://www.ijo.cn/en_publish/2018/4/20180407.pdf |
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