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HepG2 differentiated hepatic functions are retained post-thaw. (A) Urea secretion of non-frozen (grey) and freeze/thaw (blue) HepG2 cells after 1, 4 and 7 days of culture. (B) <t>CYP2C9</t> and CYP3A4 basal activity of non-frozen (grey) and freeze/thaw (blue) HepG2 cells 24 hours post-thaw. Three biological repeats were completed (ANOVA, Tukey PostHoc ns: p ≥ 0.05).
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HepG2 differentiated hepatic functions are retained post-thaw. (A) Urea secretion of non-frozen (grey) and freeze/thaw (blue) HepG2 cells after 1, 4 and 7 days of culture. (B) CYP2C9 and CYP3A4 basal activity of non-frozen (grey) and freeze/thaw (blue) HepG2 cells 24 hours post-thaw. Three biological repeats were completed (ANOVA, Tukey PostHoc ns: p ≥ 0.05).

Journal: Biomaterials Science

Article Title: Cryopreservation of assay-ready hepatocyte monolayers by chemically-induced ice nucleation: preservation of hepatic function and hepatotoxicity screening capabilities †

doi: 10.1039/d3bm01046e

Figure Lengend Snippet: HepG2 differentiated hepatic functions are retained post-thaw. (A) Urea secretion of non-frozen (grey) and freeze/thaw (blue) HepG2 cells after 1, 4 and 7 days of culture. (B) CYP2C9 and CYP3A4 basal activity of non-frozen (grey) and freeze/thaw (blue) HepG2 cells 24 hours post-thaw. Three biological repeats were completed (ANOVA, Tukey PostHoc ns: p ≥ 0.05).

Article Snippet: P450-Glo™ CYP2C9 Assay (V8792) and P450-Glo™ CYP3A4 Assay (V9002) were purchased from Promega, Wisconsin, USA.

Techniques: Activity Assay

Post-thaw primary hepatocyte function and drug response. Non-frozen (grey) hepatocytes, hepatocytes cryopreserved with 10% DMSO + IN adhered to 96-well plates (blue) and commercially supplied suspension cryopreserved hepatocytes (red) were assessed for (B) resazurin reduction (C) CYP450 activity and (D) urea secretion 24 hours post-thaw. Data is presented as an average ± SEM of 3 biological and 2 technical repeats. Drug–dose response curves were produced for non-frozen (black) and freeze/thaw adherent (blue) hepatocytes treated with (E) diclofenac (0–3.4 μM) and (F) metformin (0–77.4 mM) for 24 hours. Percentage cell viability was calculated using a resazurin reduction assay and reported ± SEM of 3 biological repeats. (ANOVA, Tukey PostHoc; ns: p ≥ 0.05, * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001).

Journal: Biomaterials Science

Article Title: Cryopreservation of assay-ready hepatocyte monolayers by chemically-induced ice nucleation: preservation of hepatic function and hepatotoxicity screening capabilities †

doi: 10.1039/d3bm01046e

Figure Lengend Snippet: Post-thaw primary hepatocyte function and drug response. Non-frozen (grey) hepatocytes, hepatocytes cryopreserved with 10% DMSO + IN adhered to 96-well plates (blue) and commercially supplied suspension cryopreserved hepatocytes (red) were assessed for (B) resazurin reduction (C) CYP450 activity and (D) urea secretion 24 hours post-thaw. Data is presented as an average ± SEM of 3 biological and 2 technical repeats. Drug–dose response curves were produced for non-frozen (black) and freeze/thaw adherent (blue) hepatocytes treated with (E) diclofenac (0–3.4 μM) and (F) metformin (0–77.4 mM) for 24 hours. Percentage cell viability was calculated using a resazurin reduction assay and reported ± SEM of 3 biological repeats. (ANOVA, Tukey PostHoc; ns: p ≥ 0.05, * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001).

Article Snippet: To determine CYP450 activity, CYP3A4 and CYP2C9 were measured using the corresponding Promega CYP450 kits (V8792 and V9002).

Techniques: Suspension, Activity Assay, Produced