3D Culture System for Liver Tissue Mimicking Hepatic Plates for Improvement of Human Hepatocyte (C3A) Function and Polarity

Table 4

Comparison between the four culture systems.

	P	S	M	H
Type of cells	C3A human hepatocytes and EA.hy926 human endothelial cells
Cell arrangement	Disorderly arrangement			Hepatic plate-like arrangement
Culture system	Mixed two-dimensional coculture/plate culture	Sandwich coculture	Mixed 3D coculture (conventional 3D culture)	Hepatic plate-mimetic 3D coculture
Cell and tissue morphology maintenance	H> M> S> P
Cell growth speed and apoptosis	P> S> M> H
Cell viability	Group H was higher than all the other groups.
Hepatocyte function and the gene expression levels	Group H exhibited significant advantages with regard to the expression of some drug-metabolizing enzymes, albumin, ammonia metabolism, and other clinically important liver function markers.
Gene expression levels of hepatocyte polarity proteins	As culture duration increased, the protein levels of occludin, DPPIV, NTCP, and CD147 in Group H were significantly higher than those in other groups.
Immunofluorescence staining of hepatocyte structural polarity proteins	Occludin, ZO-1, MRP2, and CD147 staining in Group H was much more obvious and continuous than those in other groups on days 7 and 13 as the culture duration increased.
Hepatocyte polarity ultrastructure	Group H promoted the formation of more mature bile canaliculi.
Transcriptome sequencing and analysis	Group H promoted changes in the bile secretion pathway.

H: hepatic plate-mimetic group; M: mixed 3D group; S: sandwich group; P: mixed 2D/plate group.