A semi-automated pipeline for quantifying drusen-like deposits in human induced pluripotent stem cell-derived retinal pigment epithelium cells
- Author(s)
- Hall, J; Daniszewski, M; Cheung, S; Shobhana, K; Kumar, H; Liang, HH; Beetham, H; Cho, E; Abbott, C; Hewitt, AW; Simpson, KJ; Guymer, RH; Paull, D; Pébay, A; Lidgerwood, GE;
- Details
- Publication Year 2024-06,Volume 29,Issue #3,Page 100106
- Journal Title
- SLAS Technology
- Publication Type
- Research article
- Abstract
- Age-Related Macular Degeneration (AMD) is a highly prevalent form of retinal disease amongst Western communities over 50 years of age. A hallmark of AMD pathogenesis is the accumulation of drusen underneath the retinal pigment epithelium (RPE), a biological process also observable in vitro. The accumulation of drusen has been shown to predict the progression to advanced AMD, making accurate characterisation of drusen in vitro models valuable in disease modelling and drug development. More recently, deposits above the RPE in the subretinal space, called reticular pseudodrusen (RPD) have been recognized as a sub-phenotype of AMD. While in vitro imaging techniques allow for the immunostaining of drusen-like deposits, quantification of these deposits often requires slow, low throughput manual counting of images. This further lends itself to issues including sampling biases, while ignoring critical data parameters including volume and precise localization. To overcome these issues, we developed a semi-automated pipeline for quantifying the presence of drusen-like deposits in vitro, using RPE cultures derived from patient-specific induced pluripotent stem cells (iPSCs). Using high-throughput confocal microscopy, together with three-dimensional reconstruction, we developed an imaging and analysis pipeline that quantifies the number of drusen-like deposits, and accurately and reproducibly provides the location and composition of these deposits. Extending its utility, this pipeline can determine whether the drusen-like deposits locate to the apical or basal surface of RPE cells. Here, we validate the utility of this pipeline in the quantification of drusen-like deposits in six iPSCs lines derived from patients with AMD, following their differentiation into RPE cells. This pipeline provides a valuable tool for the in vitro modelling of AMD and other retinal disease, and is amenable to mid and high throughput screenings.
- Publisher
- Elsevier
- Keywords
- Humans; *Induced Pluripotent Stem Cells/cytology; *Retinal Pigment Epithelium/pathology; *Retinal Drusen/pathology; Macular Degeneration/pathology/diagnosis; Image Processing, Computer-Assisted/methods; Microscopy, Confocal/methods; Age-related macular degeneration; Confocal microscopy; Drusen; Quantification; Reticular pseudodrusen; Retinal pigment epithelium
- Department(s)
- Laboratory Research
- Publisher's Version
- https://doi.org/10.1016/j.slast.2023.08.006
- Open Access at Publisher's Site
https://doi.org/10.1016/j.slast.2023.08.006- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2024-08-01 05:00:17
Last Modified: 2024-08-01 05:00:43