Eschenburg et al. (2018). Biomaterials. doi: 10.1016/j.biomaterials.2018.10.021
…Data presented as mean ± S.E.M (n = 2-4). Collagen isolation Collagen was isolated from rat tail tendons similar to previous descriptions [28,48,49]. Briefly, tendons were removed from rat tails (Pel-Freez Biologicals, Rogers, AR) and left to soak in 1X DPBS. The isolated collagen was then transferred into acetone for 5 minutes, followed by 70% isopropanol for an additional 5 minutes. The fibers were then…
Abstract
One of the largest challenges facing the field of tissue engineering is the incorporation of a functional vasculature, allowing effective nourishment of graft tissue beyond diffusion length scales. Here, we demonstrate a methodology for inducing the robust self-assembly of endothelial cells into stable three-dimensional perfusable networks on millimeter and centimeter length scales. Utilizing broadly accessible cell strains and reagents, we have rigorously tested a state space of cell densities (0.5-2.0 × 10<sup>6</sup> cell/mL) and collagen gel densities (2-6 mg/mL) that result in robust vascular network formation. Further, over the range of culture conditions with which we observed robust network formation, we advanced image processing algorithms and quantitative metrics to assess network connectivity, coverage, tortuosity, lumenization, and vessel diameter. These data demonstrate that decreasing collagen density produced more connected networks with higher coverage. Finally, we demonstrated that this methodology results in the formation of perfusable networks, is extensible to arbitrary geometries and centimeter scales, and results in networks that remain stable for 21 days without the need for the co-culture of supporting cells. Given the robustness and accessibility, this system is ideal for studies of tissue-scale biology, as well as future studies on the formation and remodeling of larger engineered graft tissues.