Background: Intravital microscopy in mice is widely used to study in vivo thrombus formation. Previously we have developed a 3D, tissue-engineered human arterial construct (TEAC) that replicates the primary haemostatic properties of the native artery [1].

Aims: To assess whether our TEAC can trigger the activation of the extrinsic coagulation cascade, and whether this can be used to generate a human arterial thrombosis model to replace current in vivo studies.

Methods: Medial and adventitial arterial layers were replicated by seeding human coronary artery smooth muscle cells and adventitial fibroblasts into a collagen hydrogel. A complete TEAC was created by co-culturing the medial layer with HUVEC-coated nanofibers.  Prothrombin times were measured by exposing these constructs to Platelet-poor plasma (PPP) from healthy human volunteers. A fluorogenic tissue factor assay was performed to assess tissue factor activity by incubation with factor VII and SN17a. Platelet activation was assessed using physiological flow conditions in a custom-made 3D printed flow chamber.

Results: Both the medial and adventitial layer constructs are able to trigger rapid coagulation when exposed to PPP, and have measurable tissue factor activity. Tissue factor activity in the medial layer construct was enhanced by supplementation of ascorbic acid in the culture media. Ascorbic acid supplementation of the medial construct also significantly increased platelet adhesion and aggregation on this construct under flow conditions. Incorporation of an endothelial lining atop of the media layer construct can allow us to replicate commonly used thrombosis models.

Conclusions: Our data demonstrates that our TEACs can replicate all of the major haemostatic processes of the native blood vessel, and is a viable replacement to current murine thrombosis models.
This work is supported by an NC3R- and British Heart Foundation co-funded PhD studentship 
Refs: [1] Musa et al. (2016) Tissue Eng Part C Methods 22, 691-699.

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ISTH Congress Abstracts - https://abstracts.isth.org/abstract/tissue-engineered-human-arteries-replicate-primary-and-secondary-haemostatic-functions-seen-in-vivo-a-replacement-for-mouse-arterial-thrombosis-models/