Efficient assembly of rat hepatocyte spheroids for tissue engineering applications

Florence J. Wu, Julie R. Friend, C. C. Hsiao, Michael J. Zilliox, Wen Je Ko, Frank B. Cerra, Wei Shou Hu

Research output: Contribution to journalArticlepeer-review

117 Scopus citations

Abstract

Freshly harvested primary rat hepatocytes cultivated as multicellular aggregates, or spheroids, have been observed to exhibit enhanced liver- specific function and differentiated morphology compared to cells cultured as monolayers. An efficient method of forming spheroids in spinner vessels is described. Within 24 h after inoculation, greater than 80% of inoculated cells formed spheroids. This efficiency was significantly greater than that reported previously for formation in stationary petri dishes. With a high specific oxygen uptake rate of 2.0 x 10-9 mmol O2/cell/h, the oxygen supply is critical and should be monitored for successful formation. Throughout a 6-day culture period, spheroids assembled in spinner cultures maintained a high viability and produced albumin and urea at constant rates. Transmission electron microscopy indicated extensive cell-cell contacts and tight junctions between cells within spheroids. Microvilli-lined bile canaliculus-like channels were observed in the interior of spheroids and appeared to access the exterior through pores at the outer surface. Spheroids from spinner cultures exhibited at least the level of liver-specific activity as well as similar morphology and ultrastructure compared to spheroids formed in stationary petri dishes. Hepatocytes cultured as spheroids are potentially useful three-dimensional cell systems for application in a bioartificial liver device and for studying xenobiotic drug metabolism.

Original languageEnglish (US)
Pages (from-to)404-415
Number of pages12
JournalBiotechnology and bioengineering
Volume50
Issue number4
DOIs
StatePublished - May 20 1996

Keywords

  • bioartificial liver
  • hepatocyte
  • spheroids
  • tissue engineering

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