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Human liver sinusoidal endothelial cells but not hepatocytes contain factor VIII
Author(s) -
Shahani T.,
Covens K.,
Lavend'homme R.,
Jazouli N.,
Sokal E.,
Peerlinck K.,
Jacquemin M.
Publication year - 2014
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/jth.12412
Subject(s) - flow cytometry , hepatocyte , antibody , microbiology and biotechnology , cell sorting , in vitro , cell culture , chemistry , cell , biology , immunology , biochemistry , genetics
Summary Background Although the liver is the major site of coagulation factor VIII ( FVIII ) synthesis, the type of cells producing FVIII within the liver is still unclear. Objectives To measure FVIII in extracts of primary liver sinusoidal endothelial cells ( LSEC s) and hepatocytes, thereby preventing potential bias resulting from the modifications of the cell phenotype that can take place during in vitro culture. Methods LSEC s were purified by flow cytometry cell sorting on the basis of their coexpression of Tie2 and CD 32b. The purity of the cells was controlled by RNA sequencing. FVIII activity ( FVIII :C) in extracts of purified cells was measured with a sensitive FVIII chromogenic assay, in which the specificity of the reaction is controlled by neutralization of FVIII activity with specific inhibitor antibodies. Results The FVIII:C concentration in purified LSECs ranged from 0.3 to 2.8 nU per cell. In contrast, FVIII:C was undetectable in hepatocytes. The intracellular FVIII:C concentrations are therefore at least 10–100‐fold higher in LSECs than in hepatocytes. Conclusions Our data demonstrate that LSEC s, but not hepatocytes, contain measurable amounts of FVIII :C, and suggest that the former are the main cells producing FVIII in the human liver.