May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Choroidal Neovascularization (CNV) Formation After Subretinal Injection of Homologous Retinal Pigment Epithelium (RPE) Cells and/or Polystrene Microbeads – an Experimental Mouse Model
Author Affiliations & Notes
  • X. Nie
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
  • I. Schmack
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
    Ophthalmology, Ruprecht–Karls–University, Heidelberg, Germany
  • C.L. Berglin
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
    Ophthalmology, St. Erik Eye Hospital, Karolinska Institute, Stockholm, Sweden
  • J. Wen
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
  • H. Yang
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
  • H.E. Grossniklaus
    Ophthalmology, Emory Univ Eye Clinic, Atlanta, GA
  • Footnotes
    Commercial Relationships  X. Nie, None; I. Schmack, None; C.L. Berglin, None; J. Wen, None; H. Yang, None; H.E. Grossniklaus, None.
  • Footnotes
    Support  NIH grant U10EY06360
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 3015. doi:
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      X. Nie, I. Schmack, C.L. Berglin, J. Wen, H. Yang, H.E. Grossniklaus; Choroidal Neovascularization (CNV) Formation After Subretinal Injection of Homologous Retinal Pigment Epithelium (RPE) Cells and/or Polystrene Microbeads – an Experimental Mouse Model . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3015.

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

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Abstract

Abstract: : Purpose: To establish a murine CNV–model based on subretinal injection of RPE cells and/or microbeads. To analyze the involvement of inflammatory cells and the expression of cytokines and growth factors over time. Methods: 6–8 week old wild type and lethally irradiated and GFP+/+(green fluorescent protein)–BM–reconstituted C57BL/6 mice received a subretinal injection (2µl) of a either homologous RPE cells (2x105/µl), microbeads (2x105/µl) or both in one eye. The CNV growth pattern, the presence of hematopoietic–derived inflammatory cells, and the expression of growth factors and cytokines were documented over a period of 21 days by light microscopy, immunohistochemistry, and confocal scanning laser microscopy. Thickness measurements of the CNV–complex (T), starting from the outer border of the pigmented choroidal layer up to the inner surface of the CNV, and of the retinal pigment epithelium monolayer adjacent to the lesion (P) were performed. The highest T and P values in 10 serial sections from each membrane were recorded and averaged (mean ±SD). The relative thickness of the CNV (R) in each eye was then calculated (R = T–P/P). Results: All membranes demonstrated a type 2 growth pattern with extension of the CNV into the subretinal space. A continuously increase of the CNV thickness was observed in all study groups up to day 10, followed by a gradually decrease between day 14 and 21. The CNV growth was more prominent in the PRE/microbead group (Rmax. = 31.2) compared to the two other groups (RmaxBeads = 17.8; RmaxRPE = 10.4). Inflammatory cells and various growth factors/cytokines (TGF–ß, TNF–α, TSP–1, VEGF, PEDF, TF) could be identified. The inflammatory response was less pronounced then when using heterologous RPE (abstract #1864, ARVO 2004). Conclusions: The described mouse–model is appropriate to initiate CNV growth (type 2) by subretinal injection of RPE and/or microbeads and to evaluate the different stages of CNV formation (initiation, inflammatory active, inflammatory inactive). It provides additional information regarding the inflammatory cells, cytokines and growth factors associated with CNV growth.

Keywords: age-related macular degeneration • choroid: neovascularization • retinal neovascularization 
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