Evaluation of hepatocyte phenotype maintenance in micro-aggregates.
(A) Gene expression analysis using real-time PCR. Gene expression of HNF4α, Cyp3A, Cyp1A2, E-cadherin and connexin 32 (Cx32) were determined for cells cultured on tissue culture plastic (white bars, TCP) and aggregates (black bars, AGG) at day 3, 6, 10 and 15 after isolation. The gene expression was normalized using GAPDH as stable housekeeping gene and related to freshly isolated hepatocytes (value = 1). Data are mean RQ ± SD, compared to freshly isolated, non cultured hepatocytes, n = 3. *p<0.05; **p<0.01. (B) Albumin, HNF4α, and PAS staining to visualize glycogen storage day 3 and day 10. scale bar = 50 µm. (C) Albumin secretion of plated cells and microaggregates determined by ELISA. Albumin secretion during 24 hours at day 3, 10 and 15 was evaluated for cells cultured on tissue culture plastic (white bars, TCP) and aggregates (black bars, AGG). * p<0.05, ***p<0.001