June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
An Orthotopic Transplantation Model of Retinoblastoma in Zebrafish: A Novel Gateway for Screening of Anticancer Drugs
Author Affiliations & Notes
  • Dong Hyun Jo
    Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
  • Dain Son
    Department of Microbiology and Immunology and Institute of Endemic Disease, College of Medicine, Seoul National University, Seoul, Republic of Korea
  • Yirang Na
    Department of Microbiology and Immunology and Institute of Endemic Disease, College of Medicine, Seoul National University, Seoul, Republic of Korea
  • Manyoung Jang
    Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Republic of Korea
  • Jae-Hoon Choi
    Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Republic of Korea
  • Jin Hyoung Kim
    Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
  • Young Yu
    Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
    Department of Ophthalmology, College of Medicine, Seoul National University, Seoul, Republic of Korea
  • Seung Hyeok Seok
    Department of Microbiology and Immunology and Institute of Endemic Disease, College of Medicine, Seoul National University, Seoul, Republic of Korea
  • Jeong Hun Kim
    Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
    Department of Ophthalmology, College of Medicine, Seoul National University, Seoul, Republic of Korea
  • Footnotes
    Commercial Relationships Dong Hyun Jo, None; Dain Son, None; Yirang Na, None; Manyoung Jang, None; Jae-Hoon Choi, None; Jin Hyoung Kim, None; Young Yu, None; Seung Hyeok Seok, None; Jeong Hun Kim, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1254. doi:
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      Dong Hyun Jo, Dain Son, Yirang Na, Manyoung Jang, Jae-Hoon Choi, Jin Hyoung Kim, Young Yu, Seung Hyeok Seok, Jeong Hun Kim; An Orthotopic Transplantation Model of Retinoblastoma in Zebrafish: A Novel Gateway for Screening of Anticancer Drugs. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1254.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

To establish a novel orthotopic transplantation model of retinoblastoma in zebrafish for efficient screening of anticancer drugs

 
Methods
 

We injected differential numbers (20 and 100) of retinoblastoma cells into the vitreous cavity of zebrafish at 48 hours after fertilization. Eyeballs of zebrafish were scanned daily under the confocal laser microscope. Captured images were used for quantitative analysis with the public image processing program, ImageJ. Transplanted retinoblastoma cells were isolated to perform further analyses including Western blotting for glial fibrillary acidic protein and neuron-specific enolase, reverse transcriptase for cellular retinaldehyde-binding protein, to confirm that retinoblastoma cells maintained their characteristics as tumor cells even with transplantation and isolation. To figure out the potential of this model for screening of anticancer drugs, zebrafish were treated with carboplatin and melphalan systemically after the injection of retinoblastoma cells.

 
Results
 

The degree of the tumor population estimated by the mean intensity of GFP expression was dependent on the number of retinoblastoma cells injected. The tumor population maintained stably for at least 4 days after the injection. Transplanted retinoblastoma cells maintain the proliferative potential and characteristics as retinoblastoma cells after isolation. Interestingly, systemic application of carboplatin and melphalan induced significant reduction in the tumor population, which could be quantitatively analyzed by the estimation of the mean intensity of GFP expression.

 
Conclusions
 

This orthotopic transplantation model of retinoblastoma in zebrafish is expected to be utilized for the screening of anticancer drugs for the treatment of retinoblastoma.

 
 
Retinoblastoma cells were injected into the vitreous cavity of zebrafish 48 hours after fertilization. Scale bar = 100 μm.
 
Retinoblastoma cells were injected into the vitreous cavity of zebrafish 48 hours after fertilization. Scale bar = 100 μm.
 
 
Four days after the injection, the eyeballs of zebrafish were scanned under the confocal laser microscope for further quantitative analysis. Scale bar = 100 μm.
 
Four days after the injection, the eyeballs of zebrafish were scanned under the confocal laser microscope for further quantitative analysis. Scale bar = 100 μm.
 
Keywords: 703 retinoblastoma  
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