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The effects of growth factors on the proliferation and in vitro angiogenesis of human macular inner choroidal endothelial cells

Division of Ophthalmology and Visual Sciences, University of Nottingham, Eye, Ear, Nose and Throat Centre, Queen’s Medical Centre, Nottingham, UK

Correspondence to:
Mr W M Amoaku, Division of Ophthalmology and Visual Sciences, Eye, Ear, Nose and Throat Centre, University Hospital, Queen’s Medical Centre, Nottingham NG7 2UH, UK; wma{at}nottingham.ac.uk

Aim: To investigate the effect of VEGF¹⁶⁵, FGF2, IGF-1, PDGF-AA, PDGF-BB and IL-1β on the proliferation and angiogenic tube formation of human macular inner choroidal endothelial cells (ICEC).

Methods: The proliferation of human macular ICECs after exposure to the aforementioned growth factors was determined by using both a WST-1 colorimetric assay and a cell-counting technique. The effect of growth factors on ICEC angiogenesis was assessed by sprout formation using a three-dimensional in vitro Matrigel duplex assay.

Results: Using both the WST-1 assay and a cell-counting technique, VEGF¹⁶⁵ and FGF2 both significantly increased human macular ICEC proliferation. The effect of equimolar concentrations of VEGF¹⁶⁵ and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1β on proliferation up to a growth factor concentration of 1000 pmol/l. The angiogenesis assay found a significant effect on sprout formation for VEGF¹⁶⁵ and FGF-2. Again, the effect of equimolar concentrations of VEGF¹⁶⁵ and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1β on sprout formation at 1000 pmol/l.

Conclusions: Both VEGF¹⁶⁵ and FGF2 significantly increase human macular ICEC proliferation and sprout formation in an angiogenesis assay. When present together, their effect was additive. IGF-1, PDGF-AA, PDGF-BB and IL-1β did not have any significant effect on proliferation or sprout formation in vitro. These results suggest that targeting other growth factors such as FGF2, in addition to VEGF, may be beneficial in the treatment of neovascular age-related macular degeneration.

Competing interests: None.

Ethics approval: The research had the approval of the local research ethics committee (Nottingham Q1060301).