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

Qitao Zhang; Daisy Y. Shu; Richard A. Bryan; John Y. S. Han; Gillian A. Gulette; Kin Lo; Leo A. Kim; Jason M. L. Miller

doi:10.3791/67038

Suivi à long terme des taux de consommation d’oxygène dans des cultures épithéliales pigmentaires...

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

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

Suivi à long terme des taux de consommation d’oxygène dans des cultures épithéliales pigmentaires rétiniennes hautement différenciées et polarisées

Full Text

5,591 Views

08:09 min

August 16, 2024

DOI: 10.3791/67038-v

Qitao Zhang*¹, Daisy Y. Shu*², Richard A. Bryan³, John Y. S. Han¹, Gillian A. Gulette¹, Kin Lo³, Leo A. Kim⁴, Jason M. L. Miller^1,5

¹Kellogg Eye Center,University of Michigan, Ann Arbor, ²School of Optometry and Vision Science,University of New South Wales, ³Lucid Scientific, ⁴Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology,Harvard Medical School, ⁵Cellular and Molecular Biology Program,University of Michigan, Ann Arbor

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study addresses the degeneration of retinal pigment epithelium (RPE), a key factor in vision loss, by introducing a novel device for measuring oxygen consumption rates (OCR) in RPE cultures. The Resipher device enables long-term monitoring of OCR under ideal culture conditions, allowing for extensive bioenergetic profiling.

Key Study Components

Research Area

Retinal pigment epithelium (RPE) health and function
Mitochondrial function assessment
Oxygen consumption rates (OCR)

Background

Degeneration of RPE contributes to vision loss.
Mitochondrial dysfunction is associated with RPE degeneration.
Previous OCR measurement methods were limited by non-ideal culture conditions and short monitoring time.

Methods Used

Introduction of the Resipher device for long-term OCR measurement in RPE cultures
Well-differentiated retinal pigment epithelium cell cultures
Continuous monitoring under standard cell culture conditions

Main Results

Resipher allows for measurement of OCR over weeks.
Preserves cell health while enabling detailed bioenergetic profiling.
Facilitates experimental manipulation under ideal conditions.

Conclusions

The study demonstrates an advanced method for assessing mitochondrial function in vitro.
This innovative technique is significant for RPE research and potential therapeutic interventions in vision loss.

Frequently Asked Questions

What is Retinal Pigment Epithelium (RPE)?

RPE is a layer of cells that nourishes retinal visual cells and is crucial for maintaining vision.

Why is measuring oxygen consumption rates important?

Oxygen consumption rates provide insights into mitochondrial function and overall cell metabolism.

What is the advantage of using the Resipher device?

Resipher supports long-term OCR measurements under optimal conditions, unlike previous short-term methods.

How does mitochondrial dysfunction affect vision?

Mitochondrial dysfunction can lead to RPE degeneration, ultimately causing vision loss.

Can the Resipher device be used for other cell types?

While primarily designed for RPE, it may be adaptable for other cell cultures with similar requirements.

What conditions are ideal for RPE culture?

Ideal conditions include proper nutrient supply and oxygen levels, similar to those found in vivo.

What future research can this technique enable?

This technique can facilitate studies on nutrient usage and stress response in RPE cells.

Nous présentons un nouvel appareil pour mesurer les taux de consommation d’oxygène (OCR) dans les cultures d’épithélium pigmentaire rétinien (EPR). L’appareil peut mesurer l’OCR pendant des semaines sur des EPR cultivés sur des plaques de culture cellulaire standard avec des milieux standard pendant que les plaques sont dans un incubateur de culture cellulaire standard.

La dégénérescence de l’EPR, ou épithélium pigmentaire rétinien, est à l’origine de nombreuses causes de perte de vision. Dans de nombreux cas, le dysfonctionnement mitochondrial est à l’origine de cette dégénérescence de l’EPR. Nous pouvons mesurer la fonction mitochondriale en examinant les taux de consommation d’oxygène, ou OCR.

Nous présentons ici une nouvelle méthode de mesure de l’OCR dans le RPE. Grâce aux travaux de plusieurs laboratoires, il est maintenant bien établi que l’EPR a besoin d’une activité mitochondriale pour maintenir sa santé et sa fonction. La recherche est maintenant plus axée sur les nutriments que l’EPR utilise de préférence et si cela change en cas de stress.

View the full transcript and gain access to thousands of scientific videos