title: Impaired function of bone marrow-derived endothelial progenitor cells in murine liver fibrosis.
abstract: Liver fibrosis (LF) caused by chronic liver damage has been considered as an irreversible disease. As alternative therapy for liver transplantation, there are high expectations for regenerative medicine of the liver. Bone marrow (BM)- or peripheral blood-derived stem cells, including endothelial progenitor cells (EPCs), have recently been used to treat liver cirrhosis. We investigated the biology of BM-derived EPC in a mouse model of LF. C57BL/6J mice were subcutaneously injected with carbon tetrachloride (CCl(4)) every 3 days for 90 days. Sacrificed 2 days after final injection, whole blood (WB) was collected for isolation of mononuclear cells (MNCs) and biochemical examination. Assessments of EPC in the peripheral blood and BM were performed by flow cytometry and EPC colony-forming assay, respectively, using purified MNCs and BM c-KIT(+), Sca-1(+), and Lin(-) (KSL) cells. Liver tissues underwent histological analysis with hematoxylin/eosin/Azan staining, and spleens were excised and weighed. CCl(4)-treated mice exhibited histologically bridging fibrosis, pseudolobular formation, and splenomegaly, indicating successful induction of LF. The frequency of definitive EPC-colony-forming-units (CFU) as well as total EPC-CFU at the equivalent cell number of 500 BM-KSL cells decreased significantly (p < 0.0001) in LF mice compared with control mice; no significant changes in primitive EPC-CFU occurred in LF mice. The frequency of WB-MNCs of definitive EPC-CFU decreased significantly (p < 0.01) in LF mice compared with control mice. Together, these findings indicated the existence of impaired EPC function and differentiation in BM-derived EPCs in LF mice and might be related to clinical LF.
authors.affiliation: Department of Regenerative Medicine Science, Tokai University School of Medicine, Isehara, Japan.
authors.forename: Katsuya
authors.initials: K
authors.lastname: Shirakura
authors.forename: Haruchika
authors.initials: H
authors.lastname: Masuda
authors.forename: Sang-Mo
authors.initials: SM
authors.lastname: Kwon
authors.forename: Syotaro
authors.initials: S
authors.lastname: Obi
authors.forename: Rie
authors.initials: R
authors.lastname: Ito
authors.forename: Tomoko
authors.initials: T
authors.lastname: Shizuno
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kurihara
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Mine
authors.forename: Takayuki
authors.initials: T
authors.lastname: Asahara
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000469:Alkaline Phosphatase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D001835:Body Weight; D001854:Bone Marrow Cells / Q000473:pathology*; D002454:Cell Differentiation; D019070:Cell Lineage; D003114:Colony-Forming Units Assay; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology*; D008103:Liver Cirrhosis / Q000097:blood / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009929:Organ Size; D013154:Spleen / Q000473:pathology; D013234:Stem Cells / Q000378:metabolism / Q000473:pathology*
chemical_list: D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D000469:Alkaline Phosphatase