Long-term Monitoring of Oxygen Consumption Rates in Highly Differentiated and Polarized Retinal Pigment Epithelial Cultures

Degeneration of the RPE, or retinal pigment epithelium, underlies many causes of vision loss. Mitochondrial dysfunction underlies this RPE degeneration in many cases. We can measure mitochondrial function by looking at oxygen consumption rates, or OCR.

Here we introduce a new method for measuring OCR in the RPE. Thanks to the work from multiple labs, it's now well-established, the RPE requires mitochondrial activity to maintain its health and function. Research is now more focused on what nutrients the RPE preferentially uses, and if that changes under stress.

Prior methods for assessing OCR in RPE have limitations. Either the RPE was grown in non-ideal conditions, or it could not be assessed over long periods of time. So measuring OCR in high fidelity cultures that maintain features in vivo is one of the main challenges.

The method for monitoring OCR introduced in this manuscript enables us to measure OCR over a week while manipulating the RPE under multiple experimental challenges. This is all done while the RPE is in ideal culture conditions that allows it to mimic ideal RPE in vivo. Recipher is a novel device for long-term monitoring of oxygen consumption in cell cultures, such as retinal pigment epithelium.

Unlike short-term methods like seahorse analyzer, recipher supports extended monitoring under standard conditions, enabling detailed bioenergetic profiling over weeks while preserving cell health.