Divergence in cellular markers observed in single-cell transcriptomics datasets between cultured primary trabecular...

Area of Science:

• Ophthalmology
• Genomics
• Cell Biology

Background:

• The trabecular meshwork (TM) is crucial for maintaining intraocular pressure (IOP) homeostasis.
• In vitro human trabecular meshwork (hTM) cell cultures are widely used to study TM dysfunction.
• Translating findings from in vitro hTM studies to in vivo conditions remains a significant challenge due to cell heterogeneity.

Purpose of the Study:

• To compare the transcriptomic profiles of primary hTM cells in vitro across different passages with hTM cells in tissue.
• To identify differences in cellular identity and gene expression between cultured hTM cells and those in situ.
• To establish a comprehensive transcriptomic resource for the research community to investigate TM pathobiology.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) was performed on 14 primary hTM cell culture samples (passages 1-4).
• Four of the samples were derived from individuals diagnosed with glaucoma.
• Extensive data preprocessing and quality control were applied to the scRNA-seq dataset.

Main Results:

• The study generated a comprehensive transcriptomic dataset of primary hTM cells in vitro.
• This resource allows for the study of global gene expression changes in cultured cells compared to tissue.
• The data facilitates the investigation of cellular heterogeneity and its impact on research findings.

Conclusions:

• Single-cell RNA sequencing provides critical insights into cellular identity and translatability of in vitro hTM models.
• The generated dataset serves as a public resource for advancing research on trabecular meshwork dysfunction and glaucoma.
• Understanding transcriptomic differences is key to improving the predictive value of in vitro studies for in vivo conditions.