title: A novel technique for hepatic progenitor cell isolation from normal adult rat livers.
abstract: The transplantation of hepatic progenitor cells (HPCs) is a promising alternate approach to liver transplantation for patients with end-stage liver disease. Here, we report a novel technique for HPCs isolation from normal adult rat livers (no preexposure to chemicals and no injury). HPCs were isolated from normal adult rat livers using a novel four-step collagenase perfusion method followed by density gradient centrifugation. The phenotypic properties of HPCs were characterized by morphological observation, reverse-transcription polymerase chain reaction (RT-PCR), and immunocytochemistry. The results showed that HPCs formed loose colonies and possessed a round or oval shape at culture day 3. These cells proliferated slowly and exhibited progenitor-like characteristics during the 30-day culture period. RT-PCR analysis indicated that the cultured cells were positive for several HPC-specific genes, such as albumin (ALB), alpha-fetoprotein (AFP), cytokeratin 18 (CK18), cytokeratin 19 (CK19), CD45, hepatocyte nuclear factor 1-alpha (HNF-1α), hepatocyte nuclear factor 4-alpha (HNF-4α), and Thy-1. Immunocytochemical staining showed that these cells were consistently positive for ALB, AFP, CK18, CK19, Thy-1, and OV-6. HPCs can be isolated from normal adult rat livers using a simple and effective technique involving four-step collagenase perfusion, further confirming their potential as a strong candidate for hepatocyte therapy.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Guo
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Yingyan
authors.initials: Y
authors.lastname: Lu
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Xin
authors.forename: Longyan
authors.initials: L
authors.lastname: Jiang
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Hongcui
authors.initials: H
authors.lastname: Cao
authors.forename: Yunqing
authors.initials: Y
authors.lastname: Qiu
mesh_terms: D000818:Animals; D019921:Biotinylation; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D022781:Hepatocytes / Q000166:cytology*; D007150:Immunohistochemistry / Q000379:methods; D008099:Liver / Q000473:pathology*; D008297:Male; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013234:Stem Cells / Q000166:cytology*