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Formation and differentiation of three-dimensional rat marrow stromal cell culture on microcarriers in a rotating-wall vesselUsing a high aspect ratio vessel (HARV), this study investigated the formation of 3-D rat marrow stromal cell culture on microcarriers and the expression of bone-related biochemical markers under conditions of simulated microgravity. In addition, it calculated the shear stresses imparted on the surface of microcarriers of different densities by the medium fluid in an HARV. Secondary rat marrow stromal cells were cultured on two types of microcarriers, Cytodex-3 beads and modified bioactive glass particles. Examination of cellular morphology by scanning electron microscopy revealed the presence of three-dimensional multicellular aggregates consisting of multiple cell-covered Cytodex-3 microcarriers bridged together. Mineralization was observed in the aggregates. Spherical cell-bead aggregates were observed in an HARV, while cell-bead assemblies were mostly loosely packed in a chain-like or branched structure in a cell bag. The expressions of alkaline phosphatase activity, collagen type I, and osteopontin were shown via the use of histochemical staining, immunolabeling, and confocal scanning electron microscopy. Using a numerical approach, it was found that at a given rotational speed and for a given culture medium, a larger density difference between the microcarrier and the culture medium (e.g., a modified bioactive glass particle) imparted a higher maximum shear stress on the microcarrier.
Document ID
20040120694
Acquisition Source
Johnson Space Center
Document Type
Reprint (Version printed in journal)
Authors
Qiu, Q.
(University of Pennsylvania Philadelphia [correction of Pittsburg] 19104, United States)
Ducheyne, P.
Gao, H.
Ayyaswamy, P.
Date Acquired
August 22, 2013
Publication Date
March 1, 1998
Publication Information
Publication: Tissue engineering
Volume: 4
Issue: 1
ISSN: 1076-3279
Subject Category
Aerospace Medicine
Funding Number(s)
CONTRACT_GRANT: NAG9-817
Distribution Limits
Public
Copyright
Other

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