title: Three-dimensional co-culture of hepatic progenitor cells and mesenchymal stem cells in vitro and in vivo.
abstract: BACKGROUND
  Here we co-cultured hepatic progenitor cells (HPCs) and mesenchymal stem cells (MSCs) to investigate whether the co-culture environments could increase hepatocytes form.
  
  
  METHODS
  Three-dimensional (3D) co-culture model of HPCs and MSCs was developed and morphological features of cells were continuously observed. Hepatocyte specific markers Pou5f1/Oct4, AFP, CK-18 and Alb were analyzed to confirm the differentiation of HPCs. The mRNA expression of CK-18 and Alb was analyzed by RT-PCR to investigate the influence of co-culture model to the terminal differentiation process of mature hepatocytes. The functional properties of hepatocyte-like cells were detected by continuously monitoring the albumin secretion using Gaussia luciferase assays. Scaffolds with HPCs and MSCs were implanted into nude mouse subcutaneously to set up the in vivo co-culture model.
  
  
  RESULTS
  Although two groups formed smooth spheroids and high expressed of CK-18 and Alb, hybrid spheroids had more regular structures and higher cell density. CK-18 and Alb mRNA were at a relatively higher expression level in co-culture system during the whole cultivation time (P < 0.05). Albumin secretion rates in the hybrid spheroids had been consistently higher than that in the mono-culture spheroids (P < 0.05). In vivo, the hepatocyte-like cells were consistent with the morphological features of mature hepatocytes and more well-differentiated hepatocyte-like cells were observed in the co-culture group.
  
  
  CONCLUSIONS
  HPCs and MSCs co-culture system is an efficient way to form well-differentiated hepatocyte-like cells, hence, may be helpful to the cell therapy of hepatic tissues and alleviate the problem of hepatocytes shortage.
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, 1st Youyi Road, Chongqing, 400016, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhong
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, 1st Youyi Road, Chongqing, 400016, China.
authors.forename: Juhua
authors.initials: J
authors.lastname: Gou
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, 1st Youyi Road, Chongqing, 400016, China.
authors.forename: Nian
authors.initials: N
authors.lastname: Deng
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, 1st Youyi Road, Chongqing, 400016, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Shen
authors.affiliation: Department of Surgery, Molecular Oncology Laboratory, the University of Chicago Medical Center, Chicago, Illinois, 60637.
authors.forename: Tongchuan
authors.initials: T
authors.lastname: He
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, 1st Youyi Road, Chongqing, 400016, China.
authors.forename: Bing-Qiang
authors.initials: BQ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D002454:Cell Differentiation; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods*; D022781:Hepatocytes / Q000166:cytology* / Q000637:transplantation; D007150:Immunohistochemistry; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials
keywords: AlgiMatrix™; co-culture; hepatic progenitor cells; mesenchymal stem cells; three-dimensional culture