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Enhanced liver progenitor cell survival and differentiation in vivo by spheroid implantation in a vascularized tissue engineering chamber
Yap, Kiryu Dingle, Aaron Palmer, Jason Dillon, Raminder Lokmic, Zerina Penington, Anthony Yeoh, George Morrison, Wayne Mitchell, Geraldine
Yap, Kiryu
Dingle, Aaron
Palmer, Jason
Dillon, Raminder
Lokmic, Zerina
Penington, Anthony
Yeoh, George
Morrison, Wayne
Mitchell, Geraldine
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.
Keywords
Date
2013
Type
Journal article
Journal
Biomaterials
Book
Volume
34
Issue
16
Page Range
3992-4001
Article Number
ACU Department
School of Behavioural and Health Sciences
Faculty of Health Sciences
School of Nursing, Midwifery and Paramedicine
Faculty of Health Sciences
School of Nursing, Midwifery and Paramedicine
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Source URL
Event URL
Open Access Status
License
File Access
Controlled