Journal
/
Biology
/
Real-Time Analysis of Bioenergetics in Primary Human Retinal Pigment Epithelial Cells Using High-Resolution Respirometry
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Real-Time Analysis of Bioenergetics in Primary Human Retinal Pigment Epithelial Cells Using High-Resolution Respirometry

Real-Time Analysis of Bioenergetics in Primary Human Retinal Pigment Epithelial Cells Using High-Resolution Respirometry

DOI:
10.3791/64572-v

•

09:16 min

•

February 03, 2023

•

, , , , ,
1Schepens Eye Research Institute of Mass. Eye and Ear, 2Department of Ophthalmology,Harvard Medical School

Chapters

  • 00:05Introduction
  • 00:53Culturing Human Retinal Pigment Epithelial Cells (H-RPE)
  • 02:03Assay Preparation: The Day Before Running the Assay
  • 02:43Real-Time Mito Stress Test Using the Extracellular Flux Analyzer
  • 07:26Results: Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR)
  • 08:42Conclusion

Summary

Automatic Translation

Automatic Translation

The metabolic status of human retinal pigment epithelial cells (H-RPE) reflects their health and function. Presented here is an optimized protocol for examining the real-time metabolic flux of H-RPE using high-resolution respirometry.

Tags

Keywords: Real-time BioenergeticsPrimary Human Retinal Pigment Epithelial CellsHigh-resolution RespirometryOxphosGlycolysisAMDCell CultureRespirometry InstrumentMito Stress Test Assay

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64572?article_type=v

Primary Human Bronchial Epithelial Cells Grown from Explants
check_url/64572?article_type=v

MicroRNA Expression Profiles of Human iPS Cells, Retinal Pigment Epithelium Derived From iPS, and Fetal Retinal Pigment Epithelium
check_url/64572?article_type=v

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells
check_url/64572?article_type=v

High-resolution Respirometry to Assess Mitochondrial Function in Permeabilized and Intact Cells
check_url/64572?article_type=v

Subretinal Transplantation of Human Embryonic Stem Cell Derived-retinal Pigment Epithelial Cells into a Large-eyed Model of Geographic Atrophy
check_url/64572?article_type=v

Primary Cell Cultures from the Mouse Retinal Pigment Epithelium
check_url/64572?article_type=v

Efficient Dissection and Culture of Primary Mouse Retinal Pigment Epithelial Cells
check_url/64572?article_type=v

High-Resolution Respirometry to Assess Bioenergetics in Cells and Tissues Using Chamber- and Plate-Based Respirometers
check_url/64572?article_type=v

Using Real-Time Cell Metabolic Flux Analyzer to Monitor Osteoblast Bioenergetics
check_url/64572?article_type=v

Primary Culture of Porcine Retinal Pigment Epithelial Cells

Read Article