Table 4.
Cell biology in vitro hepatic systems measuring circadian rhythmicity
| References | Cell/Tissue Type | Circadian Experimental Measure | Synchronization Method (Dose, Treatment Time) | ZT Time Period Used (Data shown in paper) | Analysis Approach |
|---|---|---|---|---|---|
| Tissue slices | |||||
| Wible et al. (129) | Liver slices from Nrf2-NULL PER2::LUC mouse (MYM and YMK strains) |
Bioluminescence of PER2-drive luciferase expression | Dexamethasone (200 nM) | 0–5 days | Data recordings excluded if bioluminescence trace did not fit a sine with >80% goodness of fit to the baseline |
| Primary cells | |||||
| Guan et al. (130) | Hepatocytes isolated from HepDKO mice | mRNA (BMAL1, Clock, Cry1, Cry2, REV-ERBα, REV-ERBβ) | None | 0, 4, 8, 12, 16, 20, 24 | JKT cycle |
| Marbach-Breitruck et al. (131) | Hepatocytes isolated from PER2::LUC mice |
Bioluminescence of PER2::luciferase expression | None | 0–180 h | Chronostar software |
| Jacobi et al. (132) | Cultured hepatocytes isolated from LBmal1KO mice | Bioenergetic assay at ZT6 and ZT18 | Not stated | 6, 18 h | Not stated |
| Liu et al. (133) | Cultured hepatocytes from LLPARDKO mice | Western blot (BMAL1) | Dexamethasone (100 nM, 1 h) | 5–35 h | Not stated |
| Wang et al. (28) | Cultured hepatocytes isolated from Lgr4m/m mice | Western blot (BMALl1) | 50% horse serum (2 h) | 0, 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48 h | Not stated for Bmal1 (JKT analysis applied for other genes of interest) |
| Torra et al. (134) | Cultured hepatocytes isolated from Sprague–Dawley rats | Western blot (REV-ERBα) | Dexamethasone (6 h) | 10, 16, 22, 28, 34, 40, 46, 52 | Mann–Whitney test comparing levels at the lowest time point (22 h) |
| Cell lines | |||||
| Dufour et al. (135) | HepG2 | mRNA (BMAL1, CLOCK, CRY1, CRY2, Per1, REV-ERBα, REV-ERBβ) | 50% horse serum (2 h) | 0, 4, 8, 12, 16, 20, 24 | Students paired t test at each time point |
| Tracey et al. (136) | HepG2 | mRNA (Nr1dp1) | 50% horse serum (2 h) | 1, 6, 11, 16, 21, 26 | Not stated |
| Lin et al. (137) | HL-7702 hepatocytes (immortalized human cell) | Western blot (PER2, REV-ERBα,) | Dexamethasone (100 nM, 2 h) | 24, 28, 32, 36, 40, 44, 48 h | Not stated |
| Wible et al. (129) | MMH-D3 hepatocytes | Bioluminescence of PER2::luciferase expression Western blot (PER2) |
Dexamethasone (200 nM) | 0, 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48 h | Data recordings excluded if bioluminescence trace did not fit a sine with >80% goodness of fit to the baseline |
| Organoids | |||||
| Leone et al. (138) | Hepanoids isolated from C57BL/6 mice | Western blot (PER2, Bmal1) | 50% horse serum | 8, 16, 24, 32, 40, 48 h | Not stated |
BMAL1, Arnt-like protein; CLOCK, circadian locomotor output cycles kaput; CRY, Cryptochrome 1/2; PER, Period 1/2/3; ZT, Zeitgeber time.