In vitro models

Toxicity – in vitro

In vitro cell-based cytotoxicity assays are useful during the early phase of drug development. These assays allow evaluation of both acute and chronic effects of test compounds.

Cornea

Human corneal epithelial cells (HCE-T¹, RIKEN, Japan) are typically used for evaluation of novel chemical entities’ or topical formulations’ cytotoxic effects. The cells can be exposed to test articles for a period of a few minutes to 3-4 days. Benzalkonium chloride (BAC), which is a typical preservative in multi-dose ophthalmic solutions, is used as positive control for HCE-T cells².

Retinal pigment epithelium (RPE)

RPE cells derived from human induced pluripotent stem cells (PCi-RPE³, Phenocell, France) introduce the key phenotypic and functional characteristics of primary human RPE cells⁴. This in vitro cell model can be used reliably to screen test articles cytotoxic effects.

Technical details

Cells

Human corneal epithelial cells (HCE-T, RIKEN, Japan)
Human induced pluripotent stem cell (iPSC)-derived retinal pigmented epithelial cells (PCi-RPE, Phenocell, France)

Cytotoxicity methods

Resazurin cell viability assay
Lactate dehydrogenase (LDH) release assay

Readouts

Cell viability

Scientific excellence in every model

Easy and cost-efficient assessment of test article-induced cytotoxicity

Screening of several test articles or concentrations at the same time

Combined use of various cytotoxic insults

References

Araki-Sasaki K, Ohashi Y, Sasabe T, Hayashi K, Watanabe H, Tano Y, Handa H. An SV40-immortalized human corneal epithelial cell line and its characterization. IOVS 1995,36:614–621. https://pubmed.ncbi.nlm.nih.gov/7534282/
Hakkarainen JJ, Reinisalo M, Ragauskas S, Seppänen A, Kaja S, Kalesnykas G. Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells. Int J Pharm. 2016;511(1):73-78. https://pubmed.ncbi.nlm.nih.gov/27374205/
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. https://pubmed.ncbi.nlm.nih.gov/23585288/
Hakkarainen JJ, Maruotti J, Seppänen A, Onteniente B, Kalesnykas G, Reinisalo M. hiPSC-derived RPE cells: Characterization of blood-retinal barrier properties and drug permeability. ARVO2016 Poster. Investigative Ophthalmology & Visual Science September 2016, Vol.57, 268.
https://iovs.arvojournals.org/article.aspx?articleid=2559157