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Induction of three-dimensional assembly of human liver cells by simulated microgravityThe establishment of long-term cultures of functional primary human liver cells (PHLC) is formidable. Developed at NASA, the Rotary Cell Culture System (RCCS) allows the creation of the unique microgravity environment of low shear force, high-mass transfer, and 3-dimensional cell culture of dissimilar cell types. The aim of our study was to establish long-term hepatocyte cultures in simulated microgravity. PHLC were harvested from human livers by collagenase perfusion and were cultured in RCCS. PHLC aggregates were readily formed and increased up to 1 cm long. The expansion of PHLC in bioreactors was further evaluated with microcarriers and biodegradable scaffolds. While microcarriers were not conducive to formation of spheroids, PHLC cultured with biodegradable scaffolds formed aggregates up to 3 cm long. Analyses of PHLC spheroids revealed tissue-like structures composed of hepatocytes, biliary epithelial cells, and/or progenitor liver cells that were arranged as bile duct-like structures along nascent vascular sprouts. Electron microscopy revealed groups of cohesive hepatocytes surrounded by complex stromal structures and reticulin fibers, bile canaliculi with multiple microvilli, and tight cellular junctions. Albumin mRNA was expressed throughout the 60-d culture. A simulated microgravity environment is conducive to maintaining long-term cultures of functional hepatocytes. This model system will assist in developing improved protocols for autologous hepatocyte transplantation, gene therapy, and liver assist devices, and facilitate studies of liver regeneration and cell-to-cell interactions that occur in vivo.
Document ID
20040141780
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Khaoustov, V. I.
(Veterans Affairs Medical Center, Baylor College of Medicine Houston, Texas 77030, United States)
Darlington, G. J.
Soriano, H. E.
Krishnan, B.
Risin, D.
Pellis, N. R.
Yoffe, B.
Date Acquired
August 22, 2013
Publication Date
October 1, 1999
Publication Information
Publication: In vitro cellular & developmental biology. Animal
Volume: 35
Issue: 9
ISSN: 1071-2690
Subject Category
Aerospace Medicine
Distribution Limits
Public
Copyright
Other
Keywords
NASA Center JSC
NASA Discipline Cell Biology

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