Table 2.
Media | timepoint | Donors (Human samples) | Animals | |||||||
---|---|---|---|---|---|---|---|---|---|---|
H1 | H26 | H27 | H29 | TX06 | TX08 | TX25 | Total | M1-M5 | ||
CTR | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 0* | 1 | 1 | 1 | 1 | 1 | 1 | 6 | 5 | |
G | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 1 | 1 | 1 | 1 | 0** | 1 | 0** | 5 | 5 | |
F | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 1 | 1 | 1 | 1 | 0** | 1 | 1 | 6 | 5 | |
GF | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 1 | 1 | 1 | 1 | 0** | 1 | 1 | 6 | 5 | |
GFI | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 1 | 1 | 1 | 1 | 0** | 1 | 1 | 6 | 5 | |
GFIO | 24 h | 0* | 1 | 1 | 1 | 1 | 1 | 0* | 5 | 5 |
48 h | 1 | 1 | 1 | 1 | 1 | 1 | 0* | 6 | 5 | |
GFIP | 24 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 |
48 h | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 7 | 5 | |
GFIPO | 24 h | 1 | 1 | 1 | 1 | 0* | 0** | 0** | 4 | 5 |
48 h | 1 | 1 | 1 | 1 | 0* | 1 | 1 | 6 | 5 | |
Total | 14 | 16 | 16 | 16 | 10 | 15 | 12 | 99 | 80 |
Sample and experimental overview. For the human study we used PCLSs from 7 human donors which have been incubated across 2 × 8 assay conditions giving a total of 112 samples. For 6 samples, the PCLSs were not suitable for RNA preparation (indicated by *). The remaining 106 samples have been processed and sequenced of which 7 samples have been identified as outliers and discarded from further analysis (indicated by **). For the mouse study we used fresh livers from five animals, which gives a total number of 16 × 5 = 80 samples. To yield enough material, three slices per sample condition have been incubated together. All mouse samples have been included. We prepared eight different media i.e. CTR: Control with 25 mM Glucose, G: 36 mM Glucose, F: 25 mM Glucose + 5 mM Fructose, GF: 36 mM Glucose + 5 mM Fructose, GFI: GF + 1 nM Insulin, GFIO: GFI + 480 μM Oleic acid, GIP: GFI + 240 μΜ Palmitic acid, GFIPO: GFI + 480 μΜ Oleic acid + 240 μΜ Palmitic acid) and cultured PCLSs from each donor for 24 h and 48 h.