Format

Send to

Choose Destination
See comment in PubMed Commons below
Circ Res. 2012 Mar 30;110(7):948-57. doi: 10.1161/CIRCRESAHA.111.263715. Epub 2012 Mar 6.

A cell-based phenotypic assay to identify cardioprotective agents.

Author information

  • 1School of Medicine, University of Rochester Medical Center, 601 Elmwood Ave, Rochester, NY 14642, USA.

Abstract

RATIONALE:

Tissue ischemia/reperfusion (IR) injury underlies several leading causes of death such as heart-attack and stroke. The lack of clinical therapies for IR injury may be partly due to the difficulty of adapting IR injury models to high-throughput screening (HTS).

OBJECTIVE:

To develop a model of IR injury that is both physiologically relevant and amenable to HTS.

METHODS AND RESULTS:

A microplate-based respirometry apparatus was used. Controlling gas flow in the plate head space, coupled with the instrument's mechanical systems, yielded a 24-well model of IR injury in which H9c2 cardiomyocytes were transiently trapped in a small volume, rendering them ischemic. After initial validation with known protective molecules, the model was used to screen a 2000-molecule library, with post-IR cell death as an end point. Po2 and pH monitoring in each well also afforded metabolic data. Ten protective, detrimental, and inert molecules from the screen were subsequently tested in a Langendorff-perfused heart model of IR injury, revealing strong correlations between the screening end point and both recovery of cardiac function (negative, r2=0.66) and infarct size (positive, r2=0.62). Relationships between the effects of added molecules on cellular bioenergetics and protection against IR injury were also studied.

CONCLUSIONS:

This novel cell-based assay can predict either protective or detrimental effects on IR injury in the intact heart. Its application may help identify therapeutic or harmful molecules.

PMID:
22394516
PMCID:
PMC3329894
DOI:
10.1161/CIRCRESAHA.111.263715
[PubMed - indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Support Center