Response of porcine hepatocytes in primary culture to plasma from severe viral hepatitis patients

Figure 1 Viability of porcine hepatocytes cultured in ♦ medium containing 10% NCS, □ 100% NP, ▲ 100% SVHP. Results were expressed as mean±SD for six samples.

Figure 2 DNA content in porcine hepatocytes grown in three different media. DNA synthesis in 100% SVHP and normal plasma was compared to that in the medium containing 10% NCS (^bP<0.01 vs 10% NCS group, by ANOVA followed by multiple comparisons). Results were expressed as mean±SD for six samples Black oblique line: 10% NCS, black small point: 100% NP; black small square: 100% SVHP.

Figure 3 TP content in porcine hepatocytes grown in three different media. TP synthesis in 100% SVHP and NP was compared to that in the medium containing 10% NCS (^aP<0.05, ^bP<0.01 vs 10% NCS group, by two-way variance analysis). Results were expressed as mean±SD for six samples. Black bar: 10% NCS; gray bar: 100% NP; white bar: 100% SVHP.

Figure 4 Leakage of LDH and AST after 5 h of culture (black bar: LDH; white bar: AST). The levels of LDH and AST in 100% SVHP were significantly higher than those in the medium containing 10% NCS. Results were expressed as mean±SD (1: t = 24.552, P = 0.001 and 2: t = 4.169, P = 0.014, compared to the10% NCS group, n = 6).

Figure 5 GSH content in porcine hepatocytes grown in three different cultures (A) (^aP<0. 05 vs 10% NCS, by two-way variance analysis, compared to 10% NCS) and CAT activity of porcine hepatocytes grown in the three different cultures (B) (^cP<0.05 vs 10% NCS by two-way variance analysis, compared to 10% NCS). Black bar: 10% NCS, gray bar: 100% NP; white bar: 100% SVHP.

Figure 6 Morphological changes of porcine hepatocytes cultured in 100% SVHP (A and B) and media containing 10% NCS (C and D). After 24 h of culture, hepatocytes in the100% SVHP group were detached from the dishes, vacuolization in the cytoplasm and deformities were more commonly observed. The membranes of cells became indistinct (A); after 48 h of culture, vacuoles were more and mainly concentrated around cell nuclei (B); after 24 h of culture, hepatocytes in the 10% NCS group constituted confluent monolayers with an intact morphology throughout the culture (C); after 48 h of culture, hepatocytes in the 10% NCS group constituted confluent monolayers with an intact morphology throughout the culture (D).