Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome

Figure 1 Fluorescence microscopy assessment of live (green-labeled) and dead (red-labeled) C3A cells at 24 h post-seeding in each group. A: Serum-free medium; B: HepatoZYME; C: DMEM/F12 with 100 mL/L FBS; D: DMEM/F12; and E: comparison of cell number from photos in each group: (group A) serum-free medium, (group B) HepatoZYME, (group C) DMEM/F12 with 100 mL/L FBS, and (group D) DMEM/F12. Data are expressed as mean ± SD (n = 3). ^aP < 0.05. FBS: Fetal bovine serum.

Figure 2 Comparison on (A) cell growth, (B) cell viability, (C) ALB secretion, (D) urea production, (E) AST leakage and (F) LDH leakage in each group. (group a) Serum-free medium, (group b) HepatoZYME, (group c) DMEM/F12 with 100 mL/L FBS, (group d) DMEM/F12. Data are expressed as mean ± SD (n = 5 in B and 3 in the others). e: Significant difference between group A and group B at the same incubation time. f: Significant difference between group A and group C at the same incubation time. α: Significant difference between group A and group D at the same incubation time. P < 0.05. AST: Aspartate transaminase; LDH: Lactate dehydrogenase; FBS: Fetal bovine serum.

Figure 3 Relative expression of genes related to hepatocyte function. A: UGT; B: GST; C: GLUL; D: G6P; E: albumin; F: CPS1; G: CYP3A4; and H: CYP2D6 in each group. (group a) Serum-free medium, (group b) HepatoZYME, (group c) DMEM/F12 with 100 mL/L FBS, (group d) DMEM/F12. Data are expressed as mean ± SD (n = 3). ^eP < 0.05, significant difference from group a at the same incubation time. UGT: Uridinediphosphate-glucuronosyl transferase; GLUL: Glutamate-ammonia ligase; CPS1: Carbamoyl phosphate synthetase I; G6P: Glucose-6-phosphatase; CYP3A4: Cytochrome P3A4; CYP2D: Cytochrome P2D6; FBS: Fetal bovine serum.

Figure 4 Fluorescence microscopy assessment of live (green-labeled) and dead (red-labeled) C3A cells 24 h post-seeding in each group. (A) serum-free medium containing 2 mg/mL sericin, (B) serum-free medium without sericin, (C) Comparison of cell number from photos in each group: (group A) Serum-free medium containing 2 mg/mL sericin, (group B) serum-free medium without sericin. Data are expressed as mean ± SD (n = 3). ^aP < 0.05, significant difference between group A and group B at the same incubation time.

Figure 5 Comparison of (A) cell growth curve and (B) cell viability curve of each group. (group a) Serum-free medium containing 2 mg/mL sericin, (group b) serum-free medium without sericin. Data are expressed as mean ± SD (n = 3 and 5, respectively). ^cP < 0.05, significant difference between group a and group b at the same incubation time.

Figure 6 Cell cycle distribution of each group. (group A) Serum-free medium containing 2 mg/mL sericin and (group B) serum-free medium without sericin. Data are expressed as mean ± SD (n = 3). The portion of cells in S phase in group A (16.21% ± 0.98%) was significantly higher than that in group B (12.61% ± 0.90%) (P = 0.009).

Figure 7 Pathway enrichment in the gene chip array.