Figure 1
From: Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
Optimized 3D collagen culture conditions promote spheroid formation and the maintenance of hepatic functions over time. (A) TPEF images of Hepoid after 8 days of culture. 3D reconstruction (xyz) of (left) 250 μm and (middle left) 100 μm depth images stack (TPEF stack) at 10× and 60× magnification, respectively; (middle right) and (right) are TPEF images taken at 20× and 60× magnification, respectively. (B) Quantification of the gene expression patterns by RT-qPCR of E-cadherin and N-cadherin in FIH (black), 2D cultures (light grey) and 3D cultured PHH (dark grey), 4 (d4), 15 (d15) and 28 days (d28) after seeding. The results are from at least three independant experiments and are expressed according to the 2D culture level (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA test, GraphPad Prism v.7.0). (C) Immunostaining of E-cadherin or N-cadherin (red), Albumin (green) and nuclei (blue) in PHH spheroids at d4, d15 and d28. (D) Immunostaining of MKL1 (pink) and nuclei (blue) in 2D and 3D cultures. Scale bar = 25 μm.
