Neonatal rat heart cells cultured in simulated microgravity |
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Authors: | Robert E Akins Nancy A Schroedl Steve R Gonda Charles R Hartzell |
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Institution: | (1) Department of Medical Cell Biology, Nemours Research Programs, Alfred I. duPont Institute, P.O. Box 269, 19899 Wilmington, Delaware;(2) Medical Sciences Division, Biotechnology Program, Johnson Space Center, NASA, 77068 Houston, Texas |
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Abstract: | Summary
In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit
gravity constrain cells to propagate, differentiate, and interact in a two-dimensional (2D) plane. Neonatal rat cardiac cells
in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by nonmyocyte cell types.
Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an
excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of
cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic
microgravity. The NASA- designed High-Aspect Ratio Vessel (HARV) bioreactors provide a low shear environment that allows cells
to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures
using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged
at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited
typical 2D organization. Cells cultured in HARVs adhered to microcarrier beads, the beads aggregated into defined clusters
containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached
to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARVs using alternative
support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV
and control cultures were similar; however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel
for the generation of tissuelike organizations of cardiac cells in vitro. |
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Keywords: | heart cells microgravity culture method HARV bioreactor tissue generation |
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