Liver epithelioid progenitor cells (LEPCs) have important roles in liver therapy because of their hepatic differentiation potency in vitro and in vivo. Despite many researches on humans, mice, and rats, equivalent progenitor cells derived from bovine are relatively rare. The purpose of our current study is to characterize bovine LEPCs, and research on the cure potency of this heteroplastic progenitor cells on mice liver fibrosis. We have used collagenase IV digesting and differential adhesion method to isolate slabstone shape, EpCAM, LGR5, NCAM1 and SOX9 positive progenitor cells from fetal Luxi bovine liver. When cultured in hepatic differentiation media containing 20 ng/ml Oncostatin M, LEPCs can differentiate into hepatocytes in vitro. After 4 weeks of intravenous tail vein injection into CCl4-injured mouse liver, LEPCs engrafted into liver parenchyma, differentiated into ALB positive hepatocytes, and could alleviate liver fibrosis through down regulating fibrosis genes-Tgfb1 and α-SMA as well as decreasing expression of collagen gene Col1a1, Col3a1, and Col4a1, and regain liver function by recovering ALT and AST. Our findings provided a useful tool for studying liver development in vitro, new cell resource for heterograft on mouse liver diseases, and a new platform for researches on immune rejection of heterogeneous cell transplantation.
Leucine rich repeat containing G protein coupled receptor 5
Liver progenitor cells
Liver epithelioid progenitor cells
Neural cell adhesion molecule 1
SRY (sex determining region Y)-box 9
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This research was supported by the National Natural Science Foundation of China (Grant Nos: 31472099; 31672404), the Agricultural Science and Technology Innovation Program (ASTIP) (cxgc-ias-01) and the project National Infrastructure of Animal Germplasm Resources (2016).
KW and HL participated in experimental designs, data acquisition and analysis, and data interpretation, as well as drafting of the manuscript. JY, CM and ZZ were involved in data acquisition and data interpretation, as well as drafting the manuscript. DZ and WG were also involved in data acquisition and data interpretation, as well as drafting the manuscript. All authors read and approved the final manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interests.
Arends B, Spee B, Schotanus BA, Roskams T, van den Ingh TS, Penning LC, Rothuizen J (2009) In vitro differentiation of liver progenitor cells derived from healthy dog livers. Stem Cells and Dev 18:351–358. doi:10.1089/scd.2008.0043CrossRefGoogle Scholar
Barker N, van Es JH, Jaks V, Kasper M, Snippert H, Toftgard R, Clevers H (2008) Very long-term self-renewal of small intestine, colon, and hair follicles from cycling Lgr5+ve stem cells. Cold Spring Harb Symp Quant Biol 73:351–356. doi:10.1101/sqb.2008.72.003CrossRefGoogle Scholar
Lee PH et al (2016) Antifibrotic activity of human placental amnion membrane-derived CD34+ mesenchymal stem/progenitor cell transplantation in mice with thioacetamide-induced liver injury. Stem Cells Transl Med 5:1473–1484. doi:10.5966/sctm.2015-0343CrossRefGoogle Scholar
Lin JS et al (2015) Hepatic differentiation of human amniotic epithelial cells and in vivo therapeutic effect on animal model of cirrhosis. J Gastroenterol Hepatol 30:1673–1682. doi:10.1111/jgh.12991CrossRefGoogle Scholar
Liu C et al (2016) Sox9 regulates self-renewal and tumorigenicity by promoting symmetrical cell division of cancer stem cells in hepatocellular carcinoma. Hepatology 64:117–129. doi:10.1002/hep.28509CrossRefGoogle Scholar
Paganelli M et al (2014) Downregulation of Sox9 expression associates with hepatogenic differentiation of human liver mesenchymal stem/progenitor cells. Stem cells Dev 23:1377–1391. doi:10.1089/scd.2013.0169CrossRefGoogle Scholar
Yoon SM, Gerasimidou D, Kuwahara R, Hytiroglou P, Yoo JE, Park YN, Theise ND (2011) Epithelial cell adhesion molecule (EpCAM) marks hepatocytes newly derived from stem/progenitor cells in humans. Hepatology 53:964–973. doi:10.1002/hep.24122CrossRefGoogle Scholar
Yovchev MI, Xue Y, Shafritz DA, Locker J, Oertel M (2014) Repopulation of the fibrotic/cirrhotic rat liver by transplanted hepatic stem/progenitor cells and mature hepatocytes. Hepatology 59:284–295. doi:10.1002/hep.26615CrossRefGoogle Scholar