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Category: Cytotoxicity

A novel assay that allows for the assessment and prediction of study compound-induced toxicity on the retinal pigment epithelium (RPE) cells in vitro.

SUMMARY

Cells

RPE cells1 (PCi-RPE1426, Phenocell, France)

Exposure

Cells are exposed to study compounds at different concentrations for selected times (e.g. 4h, 24h)

Positive control

tert-butyl hydroperoxide (tBHP) at concentrations 0-30 mM

Indicator and detection

Indicator: resazurin (7-Hydroxy-3H-phenoxazin-3-one-10-oxide) dye is irreversibly reduced to highly fluorescent metabolite resorufin (7-Hydroxy-3H-phenoxazin-3-one);

 

Detection: after 2 h exposure to resazurin, the amount of resorufin is measured at 560 nm (ex), 590 nm (em) by using Cytation 3 multi-mode reader (BioTek Instruments, Winooski, VT, USA).

Please contact us, if you would like to receive more information: info@experimentica.com

IMAGES

Characterization of hRPE cells

Expression and localization of tight junction proteins, occludin (left panel), zonula occludens 1 (ZO-1, right panel)(Hakkarainen et al., 2016) in novel RPE cells derived from human induced pluripotent stem cells (PCi-RPE1426). Scale bar 50 µm.

Transmission electron microscopy images of vertical cross sections of RPE cells showing the intracellular cell organelles, MV, microvilli; M, melanosome; N, nucleus. Scale bar 2 µm.

Novel hRPE cells (PCi-RPE1426). Scale bar 100 µm.

SELECTED PUBLICATIONS

  • Hakkarainen JJ, Maruotti J, Seppänen, Onteniente B, Kalesnykas G, Reinisalo M. hiPSC-derived RPE cells: Characterization of blood-retinal barrier properties and drug permeability. ARVO2016 Poster. Invest. Ophthalmol. Vis. Sci. 2016, Vol.57, 268.

REFERENCES

  1. Maruotti J, Wahlin K, Gorrell D, Bhutto I, Lutty G, Zack DJ. A simple and scalable process for the differentiation of retinal pigment epithelium from human pluripotent stem cells. Stem Cells Transl Med. 2013,2(5):341-354. doi: 10.5966/sctm.2012-0106.
  2. Hakkarainen JJ, Maruotti J, Seppänen, Onteniente B, Kalesnykas G, Reinisalo M. hiPSC-derived RPE cells: Characterization of blood-retinal barrier properties and drug permeability. ARVO2016 Poster. Invest. Ophthalmol. Vis. Sci. 2016, Vol.57, 268.

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