Protocols for Assessing Cell Viability in Intervertebral Disc Organ Cultures

A 31.1 MB supplementary document authored by Marcia Muerner, last updated on April 29, 2026. It provides optimized protocols for three staining methods used to assess cell viability in bovine caudal intervertebral disc organ cultures. The document compares the strengths and limitations of Calcein AM/EthD-1, LDH/EthD-1, and MTT/DAPI assays.

Use Cases

• Selecting a viability assay based on experimental time constraints, as described in the results section.
• Implementing protocol modifications for tissue penetration, based on the optimization of the Calcein AM/EthD-1 method.
• Designing appropriate positive and negative controls for viability assessment, as emphasized in the study.
• Applying insights to viability assays in other dense tissues, as suggested in the discussion.

Strengths

• Provides a direct comparison of three established staining methods (Calcein AM/EthD-1, LDH/EthD-1, MTT/DAPI).
• Includes practical guidance on protocol optimization, such as Collagenase P pre-treatment.
• Released under a CC-BY-4.0 license, facilitating open reuse.

Limitations

• Description metadata is limited; actual data quality requires manual inspection after download.
• Column-level documentation is absent; field semantics must be inferred after download.
• Row count is unknown, which may limit suitability assessment.

Provenance

Source

figshare

Collection Method

Protocol optimization and comparison study performed on bovine caudal intervertebral discs.

Freshness

Last updated 2026-04-29 05:49:32; freshness should be verified.