R2N Multicenter Oxygen Ring Trial

Reliability of Oxygen Consumption Measurements Across 9 Laboratories

Can different laboratories produce consistent oxygen consumption measurements?

This multicenter ring trial evaluates the reliability of oxygen consumption measurements in Calu-3 cell cultures as a barrier-function readout. Nine independent laboratories followed a standardized protocol.

Yes. All 9 laboratories achieved satisfactory ISO 13528 z-scores under the observed multi-center reproducibility.

Study design

Laboratories

Independent sites A to I

Batches

4 per laboratory after curation

Observations

361

Endpoints at timepoint 143

Endpoint

Oxygen %

At 24 h

Three batches (A/2, G/2, I/2) were lost entirely to oxygen instrumentation failure upstream of the present analysis and contribute missing data only.

Three key findings

The protocol works across all sites. The standardized workflow produced comparable oxygen-consumption measurements across the participating laboratories.

Between-laboratory agreement was satisfactory. The observed reproducibility was consistent with acceptable inter-laboratory performance.

Data-quality checks were essential. Instrumentation failures and missing batches were separated from the final comparative analysis.

Equivalence

Cross-site comparability

The ring trial was designed to determine whether laboratories can obtain equivalent results using the same biological system, protocol and endpoint definition.

Data Quality

Standardized protocol

All laboratories followed a common workflow to minimize procedural variation.

Curation

Failed or incomplete runs were documented and excluded from the comparative dataset where appropriate.

TEER

TEER measurements can be presented alongside oxygen-consumption data as an additional barrier-function readout and quality-control parameter.

Survey

A multicenter survey can capture practical differences between laboratories, including instrumentation, workflow handling and user experience.

Takeaways

Multicenter implementation is feasible. A harmonized protocol can support reproducible oxygen-consumption measurements across independent laboratories.