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Cytotoxic effects of alteplase, a recombinant tissue plasminogen activator, on human retinal pigment epithelial cells

Abstract

Purpose

To evaluate the cytotoxic effects of alteplase, a recombinant tissue plasminogen activator, and its additives on human retinal pigment epithelial (hRPE) cells.

Study design

Laboratory study.

Methods

We evaluated the cytotoxic effects of alteplase on human fetal RPE (hfRPE) cells, human induced pluripotent stem cell-derived RPE (hiPS-RPE), and ARPE-19 cells, as well as the cytotoxic effects of L-arginine and polysorbate 80, two additives of alteplase, on hfRPE cells. The effects of alteplase on the production of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) from hfRPE cells and the transepithelial resistance (TER) of hiPS-RPE cells were also assessed. The type of cell death induced by alteplase was investigated using ethidium homodimer III and FITC-Annexin V staining and terminal transferase deoxyuridine triphosphatase nick-end labeling.

Results

Alteplase reduced the viability of hfRPE cells significantly in a dose- and time-dependent manner. The reaction of hiPS-RPE and ARPE19 cells to alteplase was similar to that of hfRPE cells. Out of L-arginine and polysorbate 80, only treatment with L-arginine significantly reduced the viability of hfRPE cells. Alteplase (83 μg/ml, 6 h) had no significant effect on the production of VEGF and PEDF from hfRPE cells. Alteplase decreased the TER of hiPS-RPE cells in a dose- and time-dependent manner and induced necrosis as the type of cell death.

Conclusion

Alteplase can be cytotoxic to human RPE cells in a concentration- and time-dependent manner, with L-arginine being a possible causative factor.

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Acknowledgements

We thank Nobue Mukai and Kumiko Kikuchi and Shiori Ikeda for their excellent technical assistance.

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Correspondence to Yuki Morizane.

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Conflicts of interest

S. Kimura, None; Y. Morizane, None; S. Toshima, None; Y. Shiode, None; S. Doi, None; K. Takahashi, None; R.Matoba, None; Y. Kanzaki, None; F. Shiraga, None.

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Corresponding Author: Yuki Morizane

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Kimura, S., Morizane, Y., Toshima, S. et al. Cytotoxic effects of alteplase, a recombinant tissue plasminogen activator, on human retinal pigment epithelial cells. Jpn J Ophthalmol 65, 731–739 (2021). https://doi.org/10.1007/s10384-021-00848-2

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Keywords

  • Submacular hemorrhage
  • Tissue plasminogen activator
  • L-arginine
  • Human retinal pigment epithelial cell
  • Cytotoxicity