Resident endothelial cells generate hepatocytes through cell fusion in adult mouse liver

Wenjuan Pu^{1, 2, 1},
Ximeng Han^{1, 2, 1},
Mingjun Zhang^{1, 2},
Yan Li^{1, 2},
Xiuzhen Huang^{1, 2},
Lingjuan He^{1, 2},
Bin Zhou^{3, 4
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1.
State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China
3.
State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China
4.
School of Life Science and Technology, ShanghaiTech University, 100 Haike Road, Shanghai 201210, China

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Corresponding author: E-mail address: zhoubin@sibs.ac.cn (Bin Zhou)

These authors contributed equally to this work.

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These authors contributed equally to this work.

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Fig. 1. Contribution of endothelial cells (ECs) to hepatocytes through cell fusion. A, C and F: Schematic showing lineage tracing strategies (A and C) and Cdh5-DreER knock-in strategy (F). D, G and K: Schematic showing experimental procedures for tamoxifen (Tam) treatment and tissue analysis at the indicated time points after Tam treatment. B, E and H: Immunostaining for tdTomato (tdT), VE-Cad (an EC marker), and HNF4a (a hepatocyte marker) on liver sections from adult Tie2-Dre;R26-rox-tdT and Cdh5-Dre;R26-rox-tdT (B), Cdh5-CreER;R26-tdT (E), and Cdh5-DreER;R26-rox-tdT (H) mice, respectively. The percentage of tdT⁺ ECs and the cell number of tdT⁺ hepatocytes on liver sections are shown on the right. Arrowheads indicate tdT⁺ hepatocytes. 10w, 10 weeks old; +1w, 1 week after Tam treatment; +20w, 20 weeks after Tam treatment.I: Schematic showing design of dual recombinase-mediated lineage tracing of ECs and hepatocytes. J: Two possible cellular mechanisms: trans-differentiation and cell fusion. L: Fluorescence-activated cell sorting (FACS) analysis of tdT⁺ hematopoietic cells in liver, blood, and bone marrow (BM) from Alb-Cre;Cdh5-DreER;IR3 mice at 1 week after Tam treatment. M and N: Immunostaining for ZsGreen, tdT, and HNF4a on liver sections from Alb-Cre;Cdh5-DreER;IR3 mice at 1 week (M) and 12 weeks (N) of Tam treatment, respectively. Arrowheads indicate tdT⁺ZsGreen⁺ hepatocytes. O:The percentage of tdT⁺ ECs and the cell number of tdT⁺ hepatocytes on liver sections shown in M and N. Scale bars, 100 μm. Each image is representative of 3–5 individual biological samples.

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