Table 1.
Overview of read-outs used for PCLS analysis. For each enlisted technique the PCLS-related advantages, disadvantages or difficulties, analysis time (including sample preparation), and an indication of the cost per data point are given. Price range is from several euros/sample (€) to thousand euros/sample (€€€€).
| Read-Out | Advantages | Disadvantages/Difficulties | Analysis Time | Cost per Data Point |
|---|---|---|---|---|
| Viability and Cytotoxicity | ||||
| LDH leakage | Slice specific follow-up over time High throughput |
Reader required | ±1 h | € |
| ATP assay | High throughput | End point analysis only Total tissue lysis required Reader required |
±1 h | € |
| Morphology | Evaluation of liver structure | Interpretation not always straightforward Embedding technically challenging |
±3 h | € |
| Slice analysis | ||||
| qRT-PCR m(i)RNA analysis |
Evaluation of multiple m(i)RNA’s | Selective m(i)RNA measurements | ±3 h | €€ |
| Gene expression profiling | Generation of unbiased and complete mRNA profile | Need for bioinformatic skills Need for (nucleomics) infrastructure |
Several days | €€€€ |
| Western Blot | Medium sensitivity High specificity |
Low throughput High tissue input required |
1 day | €€€ |
| Hydroxyproline assay | Quantitative | High tissue input required Accurate measurement of input |
1 day | €€ |
| Immunohistochemical stainings | Can provide more detailed insight into zonation and cell type specific protein expression | Embedding technically challenging Relative low throughput Limited amount of stainings per slice |
>1 day | €€ |
| Triglyceride quantification |
Fatty/metabolic liver disease specific | Total homogenization of tissue required | ±2 h | €€ |
| GSH content | Functional | Affected by fat storage [46] | ±1 h | €€ |
| Medium analysis | ||||
| ALT, AST measurements |
Straightforward Direct indicator of liver damage |
Concentration of medium might be required/sufficient level of damage required to exceed threshold Low throughput |
Minutes | € |
| ELISA | Quantitative Multiple proteins can be tested for 1 slice |
Low throughput | Hours | €€€ |
| Protein Profiling | Generation of unbiased and complete protein profile | Need for bioinformatic skills Need for (proteomics) infrastructure |
Several days | €€€€ |