title: Enhanced liver progenitor cell survival and differentiation in vivo by spheroid implantation in a vascularized tissue engineering chamber.
abstract: Liver tissue engineering is hampered by poor implanted cell survival due to inadequate vascularization and cell-cell/cell-matrix interactions. Here, we use liver progenitor cell (LPC) spheroids to enhance cell-cell/cell-matrix interactions, with implantation into an angiogenic in vivo mouse chamber. Spheroids were generated in vitro in methylcellulose medium. Day 2 spheroids were optimal for implantation (22,407 +/-645 cells/spheroid), demonstrating maximal proliferation (Ki67 immunolabeling) and minimal apoptosis (caspase-3 immunolabelling). In vivo chambers established bilaterally on epigastric vessels of immunodeficient mice were implanted with equivalent numbers of LPCs as a cell suspension (200,000 cells), or spheroids (9 spheroids). At day 14, a trend of increased LPC survival was observed in spheroid-implanted chambers [pan-cytokeratin (panCK+) cells, p = 0.38, 2.4 fold increase)], with significantly increased differentiation [cytokeratin 18 (CK18+) cells, p < 0.002, 5.1 fold increase)] compared to cell suspension-implanted chambers. At day 45, both measures were significantly increased in spheroid-implanted chambers (panCK, p < 0.006, 16 fold increase) (CK18, p < 0.019, 6 fold increase). Hepatic acini/plates of CK18 + cells expressed hepatocyte nuclear factor 4-α and β-catenin, indicating ongoing hepatic differentiation. Spheroid cell-delivery significantly increased LPC survival and differentiation compared to conventional cell suspensions. This LPC spheroid/vascularized chamber model has clinical potential to generate three-dimensional vascularized liver tissue for liver replacement.
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia.
authors: Kiryu K
authors: KK
authors: Yap
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia.
authors: Aaron M
authors: AM
authors: Dingle
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia; Faculty of Health Sciences, Australian Catholic University, Fitzroy, Melbourne, Victoria 3065, Australia.
authors: Jason A
authors: JA
authors: Palmer
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia.
authors: Raminder S
authors: RS
authors: Dhillon
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia; University of Melbourne Department of Surgery at St Vincent's Hospital, Fitzroy, Melbourne, Victoria 3065, Australia; Murdoch Childrens Research Institute, Parkville, Victoria, Australia.
authors: Zerina
authors: Z
authors: Lokmic
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia; University of Melbourne Department of Surgery at St Vincent's Hospital, Fitzroy, Melbourne, Victoria 3065, Australia; Murdoch Childrens Research Institute, Parkville, Victoria, Australia.
authors: Anthony J
authors: AJ
authors: Penington
authors: Centre for Medical Research, Western Australian Institute for Medical Research, and School of Chemistry & Biochemistry, University of Western Australia, Perth, Australia.
authors: George C
authors: GC
authors: Yeoh
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia; University of Melbourne Department of Surgery at St Vincent's Hospital, Fitzroy, Melbourne, Victoria 3065, Australia; Faculty of Health Sciences, Australian Catholic University, Fitzroy, Melbourne, Victoria 3065, Australia.
authors: Wayne A
authors: WA
authors: Morrison
authors: O'Brien Institute, 42 Fitzroy Street, Fitzroy, Melbourne, Victoria 3065, Australia; University of Melbourne Department of Surgery at St Vincent's Hospital, Fitzroy, Melbourne, Victoria 3065, Australia; Faculty of Health Sciences, Australian Catholic University, Fitzroy, Melbourne, Victoria 3065, Australia. Electronic address: geraldine.mitchell@svhm.org.au.
authors: Geraldine M
authors: GM
authors: Mitchell
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000502:physiology*; D002454:Cell Differentiation; D002470:Cell Survival; D008099:Liver / Q000166:cytology*; D008297:Male; D051379:Mice; D016513:Mice, SCID; D018874:Spheroids, Cellular / Q000166:cytology* / Q000637:transplantation*; D013234:Stem Cells / Q000166:cytology*; D013535:Suspensions; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D013535:Suspensions