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Updated: Sep 28, 2026
Isolation, Culture, and Genetic Engineering of Mammalian Primary Pigment Epithelial Cells for Non-Viral Gene Therapy
Published on: February 26, 2021
Protective effect against ischemia and light damage of iris pigment epithelial cells transfected with the BDNF gene
Tetsuya Kano1, Toshiaki Abe, Hiroshi Tomita
1Department of Ophthalmology, Tohoku University, School of Medicine, Miyagi, Japan. t-kano@oph.med.tohoku.ac.jp
Purpose:
Brain-derived neurotrophic factor (BDNF) has been reported to protect retinal neurons against ischemia and light-induced damage. In the current study, the BDNF gene was transfected into iris pigment epithelial (IPE) cells of Long-Evans rats, and the neuroprotective ability of the IPE cells transfected with the BDNF gene against N-methyl-D-aspartate (NMDA)-induced neuroretinal cell death and against phototoxic damage was examined.
Methods:
The level of BDNF mRNA and protein expressed in the transfected cells was determined by reverse transcription-polymerase chain reaction (RT-PCR) and by sandwich enzyme-linked immunosorbent assay (ELISA). The neuroprotective effects were determined by culturing BDNF gene-transfected IPE cells or nontransfected cells with neuroretinal cells in the presence of NMDA. The neuroprotective activity was also evaluated for the damage induced by constant exposure to light on the photoreceptors by transplanting BDNF gene-transfected IPE cells into the subretinal region of the superior half of the eye.
Results:
BDNF gene-transfected IPE cells expressed higher levels of BDNF mRNA and protein than did nontransfected IPE cells. A significant increase in the protection against NMDA was observed in the neuroretinal cells cultured with BDNF-transfected IPE cells than in those cultured with nontransfected IPE cells (P = 0.0029) or with nontreated cells (P = 0.0010). The effect was partially attenuated by the addition of anti-BDNF antibody. Significant photoreceptor cell protection against injury from constant light was also observed by the subretinal transplantation of BDNF-transfected IPE cells when compared with those receiving transplants of nontransfected cells or vehicle injection.
Conclusions:
BDNF-transfected IPE cells demonstrated a neuronal rescue effect in both in vitro and in vivo experiments. IPE cells may be a potential source for autologous cell transplantation for some retinal diseases, and the transfection of the genes of neurotrophic factors into the transplanted cell may be a useful tool for delivering these factors to the retina.
