ASGSB 1998 Annual Meeting Abstracts

[78]
CHARACTERIZATION OF 3-DIMENSIONAL VASCULAR CELL CO-CULTURES MAINTAINED IN THE ROTATING BIOREACTOR.   D. Ellerson, G.L. Sanford, S.A. Harris-Hooker and C.D. Melhado, Space Medicine & Life Sciences Research Center, Morehouse School of Medicine, Atlanta, GA.

Vascular remodeling is a complex set of events involving endothelial cell injury and/or dysfunction that results in intimal/medial thickening. Although this area has received significant attention, the cellular and molecular mechanisms of vascular remodeling are not completely understood. The development of 3-D co-culture models of the blood vessel will provide a unique opportunity to conduct mechanistic studies into vascular remodeling. We characterized the 3-D growth of endothelial (EC) and smooth muscle (SMC) cells, alone and in co-culture, using the NASA horizontally rotating bioreactor (HRB). Cells were continuously cultured on cytodex-3 microcarriers for up to 30 days (HRB and SF) and were processed for scanning electron microscopy examination, immunocytochemical assessment of phenotypic marker. Controls were maintained in spinner flasks (SF) over the same period. In both systems, microcarriers and cells remain uniformly suspended in the fluid. We found that both EC and SMC grew at a slower rate in the HRB than in the SF. All cultures grew as 3-D aggregates after 14 days. These cultures were positive for the von Willebrand factor (EC) and alpha actin (SMC). The glucose consumption were monitored as an index of cell growth. The cross section for Transmission Electron Micrographs demonstrate the ultrastructural characteristics of SMC and EC. With the large aggregates formed by co-cultures, the surface EC appear to be invaginating, and after 30 days, tube-like structures can be seen in the interior of aggregates. These results suggest that in the HRB, vascular cells spontaneously form 3-D capillary like structures. Hence such cultures may provide a unique model for mechanistic studies of vascular remodeling and angiogenesis.
(Supported by grants from NASA: NAG9-852 & NCCW-0083 and NIH/RCMI (3G12 RR03034).

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