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. 2021 Nov 8;9:738081. doi: 10.3389/fbioe.2021.738081

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Copyright © 2021 Liu, Zhou, Chen, Lv, Huang, Peng, Wu, Chen, Tang, Guo, Wang, Zhang, Liu, Yang, Yu and Yan.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Establishment of a co-culture system using HepLPCs and HUVECs. (A) Representative bright-field images of HepLPCs in expandable monolayer HepLPCs, HepLPCs spheroids, vHepLPC spheroids, and differentiated monolayer HepLPCs. (B) Gene expression was analyzed by qRT-PCR. Hepatic functional markers (ALB, HNF4A) were detected in four groups. Data are normalized to the expandable monolayer HepLPCs group. (C) Fluorescence morphology of HepLPC spheroids on carriers. (D–F) Periodic acid-Schiff (PAS) staining, oil red staining, and ICG staining of HepLPCs (upper panel) and vHepLPC spheroids (lower panel). (G) The expression of ALB was determined by immunofluorescent staining in vHepLPCs. Nuclei were counterstained with DAPI. Data are shown as mean ± SEM. **p < 0.01. Scale bars, 100 µm.