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. 2025 Aug 12;12:1562641. doi: 10.3389/fvets.2025.1562641

Table 1.

THY toxicity in different animal cell lines.

Cell type Cell culture/cell line and species of origin Assay Exposure Findings References Year
Enterocytes Canine BMSC (primary cell culture) and canine BMSC differentiated into ELC cells (primary cell culture) Alamar blue THY: 4.66–2,529.67 μM THEO (THY: 40%): 11.32–6,113.89 μM LC50 for THY is 13.32 ± 0.67 μM (BMSC) and 13.32 ± 5.33 μM (ELC) LC50 for THEO is 533.06 ± 19.99 μM (BMSC), 393.04 ± 6.66 μM (ELC) (30) 2015
IPEC-J2 cells (cell line; pig) WST-1 THY: 10–200 μM THY at <100 μM shows no significant effect on viability; at >100 μM, it significantly reduces viability (25) 2019
Trypan blue staining THY-CIN blend: 0.0067–6656.37 μM; 48 h High viability (>90%) is observed for concentrations up to 6.66 μM (31) 2020
Hepatocytes Porcine hepatocytes (primary cell culture) Not reported THEO from three different Thymus species (THY: 38.5%, 49.10% and 56.02%): >2,663.44 μM No toxicity is detected for any of the EO tested (32) 2013
Canine hepatocytes (primary cell culture) Alamar blue THY and THEO (THY: 40%): 0.0665–6.665 μM LC50 for THY is 0.33 ± 0.07 μM
LC50 for THEO is 1.20 ± 0.16 μM
(33) 2015
LMH cells (cell line; chicken) Trypan blue staining THY-CIN blend (75% 25%); 0.0067 μM a 667 μM; 48 h High viability (>90%) is observed up to a maximum of 0.67 μM (34) 2020
Renal Cells Vero (cell line; African green monkey) XTT THY: 25–400 μM; 4 days CC50: 300 μM (35) 2012
MDCK (cell line; dog) BAPTA/AM chelation) THY: 300–500 μM Cytotoxicity not prevented by Ca2+ chelation with BAPTA/AM (36) 2014
MDCK (cell line; dog) Alamar blue THY: 0.3–0.06 mg/ml and THEO (THY: 40%): 0.18–0.03 mg/ml THY is more toxic (LC50 = 0.13 ± 0.002 mg/ml) than THEO (LC50 = 0.16 ± 0.01 mg/ml) (37) 2015
FRhK-4 (cell line; monkey) Visual inspection under optical microscope THY: 0.1–2%. Cytotoxicity is observed at THY 2%. (38) 2015
CRFK (cell line; cat) XTT and visual inspection under optical microscope THEO (THY: 47%): 8,900, 4,450, 2,225, 1,112.5, 556.25, 278.12, 139.06, 69.53 μg/mL CC20 for THEO: 194.98 μg/mL (39) 2024
Splenocytes Rat splenocytes (primary cell culture) MTT THY: 0.5–500 μM IC50: 362 μM (40) 2013
Macrophages RAW 264.7 cells (cell line; mice) MTT THY and derivatives (acetyl-THY and benzoyl-THY): 0–100 μg/mL; 48 h THY at 100 μg/mL reduces survival to 36.5%; benzoyl-THY at 100 μg/mL decreases macrophage survival to 63.6%, and acetyl-THY at 100 μg/mL shows >100% survival (3) 2014
Visual inspection under optical microscope THY: 0.1–2%. Cytotoxicity is observed at THY 2%. (38) 2015
MTT THY: 10–40 μg/mL; 24 h No cytotoxicity is observed at tested doses (41) 2017
MTT THY: 25–400 μM No cytotoxicity is observed at tested doses (42) 2018
RAW 264.7 cells (cell line; mice) and bone marrow macrophages (primary cell culture) MTT THY: 10–40 μM; 24 h No cytotoxicity is observed at tested doses (26)
Leukocytes Mice peritoneal leukocytes (primary cell culture) MTT THY (1.5–150 μg/mL) Viability remains >83% at all tested concentrations (43) 2012
Monocytes HD11 monocytes (cell line; chicken) Trypan blue staining THY-CIN blend (75% 25%; 1–100,000 ng/ml; 48 h High viability (>90%) is observed (34) 2020
Peripheral blood mononuclear cells Cattle PBMC (primary cell culture) MTT THY:1 and 10 Mm No cytotoxicity is observed at tested doses (44) 2023
Mammary epithelial cells Mouse mammary epithelial cells (primary cell culture) MTT THY: 10–40 μg/mL; 24 h No cytotoxicity is observed at tested doses (24) 2014
Bovine mammary epithelial cells (primary cell culture) CCK-8 THY: 16–64 mg/mL; 48 h No cytotoxicity is observed at tested doses (45)
Fibroblasts NIH-3T3 fibroblasts (cell line; mice) MTT Enriched cellulose hydrogel with THY: 0.25–1% Viability decreases slightly with increasing concentration (98.84 to 88.81%); overall, low toxicity is observed (46) 2018