Abstract
An emerging challenge in tissue engineering biomimetic models is recapitulating the physiological complexity associated with real tissues. Recently, our laboratory introduced the rat mesentery culture model as an ex vivo experimental platform for investigating the multi-cellular dynamics involved in angiogenesis within an intact microvascular network using time-lapse imaging. A critical question remains whether the vessels maintain their functionality. The objective of this study was to determine whether vascular smooth muscle cells in cultured microvascular networks maintain the ability to constrict. Adult rat mesenteric tissues were harvested and cultured for three days in either MEM or MEM plus 10% serum. On Day 0 and Day 3 live microvascular networks were visualized with FITC conjugated BSI-lectin labeling and arteriole diameters were compared before and five minutes after topical exposure to vasoconstrictors (50 mM KCl and 20 nM Endothelin-1). Arterioles displayed a vasoconstriction response to KCl and endothelin for each experimental group. However, the Day 3 serum cultured networks were angiogenic, characterized by increased vessel density, and displayed a decreased vasoconstriction response compared to Day 0 networks. The results support the physiological relevance of the rat mesentery culture model as a biomimetic tool for investigating microvascular growth and function ex vivo.
Highlights
Tissue culture approaches offer alternative top-down strategies to maintain the complexity of tissues ex vivo[7,8,9]
Vascular smooth muscle cells maintain the ability to constrict in cultured ex vivo rat mesenteric tissues
The main contribution of this study is the establishment of the rat mesentery culture model as an ex vivo experimental platform in which smooth muscle cells (SMCs) remain functional
Summary
Tissue culture approaches offer alternative top-down strategies to maintain the complexity of tissues ex vivo[7,8,9]. As an example of a top-down tissue culture strategy, our laboratory recently introduced the rat mesentery culture model to investigate cell-cell interactions during microvascular remodeling in viable, intact microvascular networks[10]. The objective of this study was to determine if the smooth muscle cells (SMCs) within the rat mesenteric microvascular network maintain the ability to constrict during culture. The findings from this study provide new information regarding the physiological relevance of the rat mesentery culture model as a tool to investigate microvascular growth and function within an intact microvascular network and support the potential of top-down tissue culture methods for biomimetic tissue engineering applications. The identification of impaired vasoconstriction in networks characterized by angiogenesis based on our culture model data and chronic in vivo data further support a novel discovery for angiogenesis research. The observation of impaired function in remodeling vessels is new or at least underappreciated and motivates a platform for future investigation
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