Ocular Toxicity Evaluation by Human Retinal Pigment Epithelial (RPE) Cells

Creative Biolabs leverages cutting-edge insights and expert teams to provide ocular toxicity evaluation service by human RPE cells.

RPE Cells Overview

Anatomical position
RPE cells are located in the posterior part of the retina, adjacent to photoreceptor cells (e.g., cone and rod cells), and their dorsal surface is immediately adjacent to the choroid, which is the cell layer where ocular toxicity events are often manifested and are suitable as a modeling tool for evaluation.
Morphological characteristics
RPE cells present a hexagonal cobblestone morphology with pigmentation and multiple microvilli that increase the surface area and help to adsorb and stabilize photoreceptor outer segments above the optic cones and optic rod cells.
Function
- Protect and support cone and rod cells by providing nutrients, removing waste products, and regulating electrolyte balance.
- Enriched with melanin to absorb and block excess light to prevent it from reflecting back onto the cone and rod cells, reducing light interference and scattering.
- Involved in the metabolism and regeneration of retinol, maintaining visual light signaling.

Maturation of RPE cells

Beginning

Beginning with human iPSCs, it is possible to allow for combinatorial evaluation with sources of the same genetic background as the retinal organoid.

Differentiation

iPSCs selectively differentiate into retinal progenitor cells, which then receive stimulation by the active Wnt signaling pathway, as well as regulatory transcription factors such as OTX2 and MITF, to further differentiate into RPE progenitors. Final differentiation into RPE cells after induced accumulation of specific proteins and precipitated pigments.

Cultivation conditions

Specific media and growth factors are used to mimic environmental conditions in vivo. Typical media include DMEM/F12 or RPMI 1640 and contain specific additives.

Validating RPEs

Confirmation of RPEs characteristics and functions by molecular biology techniques such as immunofluorescence staining and PCR. For example, detecting the expression of specific markers (BEST1, MITF, RPE65, etc.).

Extensive Tests for Evaluation

Cell survival tests

For example, MTT test, fluorescent staining of cells, and ATP assay to determine toxicity by comparing the survival rates of treated and control groups.

Cell damage tests

Detecting biomarkers of cellular damage, such as cell membrane integrity assay, cell polarity, phagocytic activity of the outer segments of the photoreceptors, lysosomal pH, and enzyme activities, to assess the effect of compounds on RPE cell membrane integrity and cellular damage.

Oxidative stress tests

Measuring the antioxidant enzyme activities (e.g., superoxide dismutase, glutathione peroxidase) or measurement of indicators of oxidative stress (e.g., ROS production, changes in antioxidant content).

Cellular inflammation tests

Measuring the expression level or activity of inflammation-related mediators to assess the effect of the evaluated compounds on the inflammatory response of RPE cells.

Data analysis

Curve plotting (e.g., dose-response curves presenting the relationship between compound concentration and cell survival), IC50 calculation, etc.

In vitro characterization of human embryonic stem cell (hESC)-derived retinal pigment epithelium (RPE) cells. Fig 1. Characterization of hESC-derived RPE cells in vitro.¹

More 3D Biology Based Ocular Toxicity Evaluation Services

Creative Biolabs provides high-quality ocular toxicity evaluation with RPE cells to help advance scientists' research on the effects of eye-related drugs. Please contact us to get a customized solution.

Reference

Dong, Chunxiao et al. "Ex vivo cultivated retinal pigment epithelial cell transplantation for the treatment of rabbit corneal endothelial dysfunction." Eye and vision (London). 10,1 (2023): 34. Distributed under Open Access license CC BY 4.0, without modification.

Research Model

3D Biology Based Biomarker Discovery