Effects of collagen gel configuration on behavior of vascular smooth muscle cells in vitro: Association with vascular morphogenesis |
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Authors: | Jian Song Barbara E Rolfe Ian P Hayward Gordon R Campbell Julie H Campbell |
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Institution: | (1) Center for Research in Vascular Biology, Department of Anatomical Sciences, University of Queensland, 4072 Brisbane, Queensland, Australia |
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Abstract: | Summary The growth, behavior, and contractile protein expression of rabbit aortic smooth muscle cells (SMC) grown on, between layers,
or within a collagen gel was investigated by confocal laser scanning fluorescence microscopy and Western analysis. SMC grown
on collagen gel behaved similarly to those on conventional culture dishes. However, when a second layer of collagen was overlaid,
cells underwent an elongated quiescent phase before onset of proliferation and a more than threefold lower logarithmic growth
rate was observed. These cells self-organized into a network with ring-like structures. With increasing culture time, some
of the rings developed into funnel-like, incomplete or complete tubular structures. If a tubular template preexisted within
the gel, the SMC established a cylinder-shaped tube with several circularly arranged muscular layers (similar to an artery
wall). This behavior mimicked endothelial cells during angiogenesis in vitro. A similar phenomenon occurred in cultures in
which SMC were randomly mixed in a collagen gel, but here their behavior and morphology varied with their position within
the gel. Western blot analysis showed that the SMC differentiation marker, smooth muscle myosin heavy chain-2 (SM-2), rapidly
decreased, disappearing by day 10 in SMC grown on collagen, but was still detectable until day 25 in cells cultured between
or within the same gel. These findings indicate that like endothelial cells, vascular SMC can display blood vessel formation
behavior in vitro when an appropriate three-dimensional matrix environment is provided to keep them in a relatively higher-differentiated
and low-proliferative state. |
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Keywords: | extracellular matrix cell culture vascular development cell growth differentiation |
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