June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Comparison of the laser-induced choroidal neovascularization model pathology in C57BL/6J mice from different vendors
Author Affiliations & Notes
  • Maria Vähätupa
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Giedrius Kalesnykas
    R&D Division, Experimentica Ltd., Kuopio, Finland
    R&D Division, Experimentica Ltd., Vilnius, Lithuania
  • Ville Jokinen
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Teemu Nevalainen
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Leena Tähtivaara
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Rubina Thapa
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Heidi Koskenniemi
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Anni Tenhunen
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Birgitta Lappeteläinen
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Anne Mari Haapaniemi
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Nerija Kvietkauskiene
    R&D Division, Experimentica Ltd., Vilnius, Lithuania
  • Jonas Jarutis
    R&D Division, Experimentica Ltd., Vilnius, Lithuania
  • Simas Bijeikis
    R&D Division, Experimentica Ltd., Vilnius, Lithuania
  • Simon Kaja
    R&D Division, Experimentica Ltd., Forest Park, Illinois, United States
    Ophthalmology and Molecular Pharmacology & Neuroscience, Loyola University, Chicago, Illinois, United States
  • Marc Cerrada-Gimenez
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Footnotes
    Commercial Relationships   Maria Vähätupa Experimentica Ltd., Code E (Employment); Giedrius Kalesnykas Experimentica Ltd, Code E (Employment), Experimentica Ltd, Code I (Personal Financial Interest), Experimentica Ltd, Code O (Owner); Ville Jokinen Experimentica Ltd, Code E (Employment); Teemu Nevalainen Experimentica Ltd., Code E (Employment); Leena Tähtivaara Experimentica Ltd., Code E (Employment); Rubina Thapa Experimentica Ltd., Code E (Employment); Heidi Koskenniemi Experimentica Ltd., Code E (Employment); Anni Tenhunen Experimentica Ltd., Code E (Employment); Birgitta Lappeteläinen Experimentica Ltd., Code E (Employment); Anne Mari Haapaniemi Experimentica Ltd., Code E (Employment); Nerija Kvietkauskiene Experimentica Ltd., Code E (Employment); Jonas Jarutis Experimentica Ltd., Code E (Employment); Simas Bijeikis Experimentica Ltd., Code E (Employment); Simon Kaja Experimentica Ltd., Code C (Consultant/Contractor), Experimentica Ltd., Code E (Employment), Experimentica Ltd., K&P Scientific LLC, Code F (Financial Support), Experimentica Ltd., K&P Scientific LLC, Code I (Personal Financial Interest), eyeNOS Inc., Code P (Patent), Experimentica Ltd., K&P Scientific LLC, Code R (Recipient), Experimentica Ltd., K&P Scientific LLC, Code S (non-remunerative); Marc Cerrada-Gimenez Experimentica Ltd., Code E (Employment)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3032 – F0403. doi:
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      Maria Vähätupa, Giedrius Kalesnykas, Ville Jokinen, Teemu Nevalainen, Leena Tähtivaara, Rubina Thapa, Heidi Koskenniemi, Anni Tenhunen, Birgitta Lappeteläinen, Anne Mari Haapaniemi, Nerija Kvietkauskiene, Jonas Jarutis, Simas Bijeikis, Simon Kaja, Marc Cerrada-Gimenez; Comparison of the laser-induced choroidal neovascularization model pathology in C57BL/6J mice from different vendors. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3032 – F0403.

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

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Abstract

Purpose : The mouse laser-induced choroidal neovascularization (CNV) model is the most popular experimental model to mimic the wet form of age-related macular degeneration (AMD). In this study we aimed to compare the differences in CNV pathology between C57BL/6J mice from different vendors.

Methods : 8-week-old male C57BL/6JRj mice from Janvier Labs (France) and C57BL/6JRccHsd from Envigo (Netherlands) were used in the study. CNV was induced by using a 532 nm laser and the development was monitored in vivo by fluorescein angiography (FA) and spectral domain optical coherence tomography (SD-OCT) on days 0, 3, 7 and 14. The lesions were qualitatively graded from FA images for vascular leakage (non-leaky or leaky). FA scans were analysed by a proprietary algorithm, where the neural network was trained to recognize and quantify CNV lesions using transfer learning approach. Choroidal flat mounts (D7 n = 5-6/group or D14 n = 6/group) were immunolabelled with isolectin B4, anti-iba1 and anti-collagen I antibodies, scanned using Leica THUNDER 3D microscope and manually analysed using FIJI/ImageJ (NIH). The data were statistically analysed with Mixed-effects model (FA) or Mann-Whitney U test (histology).

Results : There was no difference in the CNV grading (% of leaky lesions) in Janvier mice compared to Envigo (p=0.96). While absolute values of FA leakage area were overall lower in Janvier mice in all time points, this study was not sufficiently powered to detect that difference (area in mm2, mean ± SD, Janvier vs. Envigo: 0.04±0.042 vs. 0.056±0.054 on D3; 0.014±0.028 vs. 0.043±0.092 on D7; 0.008±0.015 vs. 0.035±0.067 on D14). Isolectin area was larger in Envigo mice lesions on D7 (p <0.05) and on D14 (p <0.0001) as compared to Janvier mice. Iba1 area was bigger in Envigo mice as compared to Janvier mice on D7 (p <0.001). There was a trend towards bigger Collagen I area in Envigo animals compared to Janvier mice on D7 (p =0.077).

Conclusions : Envigo C57BL/6JRccHsd mice show more severe CNV pathology than Janvier C57BL/6JRj mice. These data suggest strain-specific differences in neovascular and fibrotic processes after laser CNV induction. These differences should be considered when selecting animal strains for IND-enabling studies in wet AMD.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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