TEMPORARY REMOVAL: CD271 identifies functional human hepatic stellate cells, which localize in peri-sinusoidal and portal areas in liver after partial hepatectomy.
Hepatic stellate cells (HSCs) are liver-resident mesenchymal cells involved in essential processes in the liver. However, knowledge concerning these cells in human livers is limited because of the lack of a simple isolation method.



We isolated fetal and adult human liver cells by immunomagnetic beads coated with antibodies to a mesenchymal stromal cell marker (CD271) to enrich a population of HSCs. The cells were characterized by cell cultivation, immunocytochemistry, flow cytometry, reverse-transcription polymerase chain reaction and immunohistochemistry. Cells were injected into nude mice after partial hepatectomy to study in vivo localization of the cells.



In vitro, CD271(+) cells were lipid-containing cells expressing several HSC markers: the glial fibrillary acidic protein, desmin, vimentin and α-smooth muscle actin but negative for CK8, albumin and hepatocyte antigen. The cells produced several inflammatory cytokines such as interleukin (IL)-6, IL-1A, IL-1B and IL-8 and matrix metalloproteinases MMP-1 and MMP-3 and inhibitors TIMP-1 and TIMP-2. In vivo, fetal CD271(+) cells were found in the peri-sinusoidal space and around portal vessels, whereas adult CD271(+) cells were found mainly in the portal connective tissue and in the walls of the portal vessels, which co-localized with α-smooth muscle actin or desmin. CD271(-) cells did not show this pattern of distribution in the liver parenchyma.



The described protocol establishes a method for isolation of mesenchymal cell precursors for hepatic stellate cells, portal fibroblasts and vascular smooth muscle cells. These cells provide a novel culture system to study human hepatic fibrogenesis, gene expression and transcription factors controlling HSC regulation.
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden.
Pradeep B
PB
Patil
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden.
Meghnad
M
Joshi
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden.
Vijay Kumar
VK
Kuna
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden.
Bo
B
Xu
Gynecology and Obstetrics at Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Liza
L
Johannesson
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden.
Michael
M
Olausson
Sahlgrenska University Hospital, Laboratory for Transplantation and Regenerative Medicine, University of Gothenburg, Gothenburg, Sweden. Electronic address: suchitra.holgersson@gu.se.
Suchitra
S
Sumitran-Holgersson
D000818:Animals; D016207:Cytokines / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006498:Hepatectomy; D055166:Hepatic Stellate Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000601:surgery; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism*
D016207:Cytokines; D005904:Glial Fibrillary Acidic Protein; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D017475:Receptors, Nerve Growth Factor
Ito cells; peri-sinusoidal cells; stellate cells