Long-term viability and proliferation of alginate-encapsulated 3-D HepG2 aggregates formed in an ultrasound trap.

D. Bazou*, W. T. Coakley, A. J. Hayes, S. K. Jackson

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

We report proof of principle here of a gel encapsulation technique that departs from the minimum surface area to volume restriction of spherical microcapsules and allows gelation of preformed high-density (>or=2x10(4) cells/aggregate) 3-D HepG2 cell aggregates. The process involves forming a discoid 3-D cell aggregate in an ultrasound standing wave trap (USWT), which is subsequently recovered and encapsulated in alginate/CaCl2 hydrogel. The size of the ultrasound-formed aggregates was dependent upon the initial cell concentration, and was in the range of 0.4-2.6 mm in diameter (for cell concentrations ranging between 10(4) and 5x10(6)/ml). At low cell concentrations (<or=5x10(5)/ml), aggregates were 2-D, while at concentrations of >or=10(6)/ml, 3-D aggregates were generated. Cells in non- and encapsulated 3-D HepG2 aggregates remained 70-80% viable over 10 days in culture. The proliferative activity of the aggregates resulted in the doubling of the aggregate cell number and a subsequent increase in the aggregate thickness, while albumin secretion levels in encapsulated aggregates was 4.5 times higher compared to non-encapsulated, control aggregates. The results reported here suggest that the ultrasound trap can provide an alternative, novel approach of hydrogel cell encapsulation and thus rapidly (within 5 min) produce in vitro models for hepatocyte functional studies (for example, toxicity studies particularly if primary hepatocytes are used) in a tissue-mimetic manner.
Original languageEnglish
Pages (from-to)1321-1331
Number of pages0
JournalToxicol In Vitro
Volume22
Issue number5
DOIs
Publication statusPublished - Aug 2008

Keywords

  • Actins
  • Albumins
  • Alginates
  • Biocompatible Materials
  • Calcium Chloride
  • Carcinoma
  • Hepatocellular
  • Cell Aggregation
  • Cell Death
  • Cell Proliferation
  • Cell Survival
  • Hepatocytes
  • Humans
  • Hydrogel
  • Polyethylene Glycol Dimethacrylate
  • Ultrasonics

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