However, the archetypal approach of using biodegradable scaffolds to provide initial strength for the newly constructed vessels raises concerns about foreign body reaction, inflammation and infection due to bacterial colonization,. Įarly approaches towards blood vessel tissue engineering are termed “top down” approaches and are based on seeding cells on porous scaffolds or embedding cells in hydrogels to support the cells and achieve the formation of tubular tissue engineered constructs. To address the current limitations in treating small diameter blood vessel disease, researchers in the medical community have strived to develop a tissue engineered small diameter artery for more than two decades,. Similarly, the use of synthetic grafts in coronary artery (and other small diameter arteries) augmentation is clinically unsuccessful due to loss of patency which occurs due to thrombus accumulation on the lumen of the graft. However, this strategy is suboptimal due to second site morbidity, a limited supply of autografts, and loss of patency, ,. The current gold standard in coronary artery bypass surgery is a blood vessel autograft where a vessel explanted from the patient is attached to the diseased artery in order to reroute blood around the obstruction and restore blood flow to the heart. This method improves cell sheet handling, results in rapid circumferential alignment of smooth muscle cells which immediately express contractile genes, and introduction of an analogue to small diameter blood vessel IEL. Upon cooling to room temperature, the scaffold, its adherent cells, and the remaining cell sheet detached and were collected on a mandrel to generating tubular constructs with circumferentially aligned smooth muscle cells which possess contractile gene expression and a single layer of electrospun scaffold as an analogue to a small diameter blood vessel's internal elastic lamina (IEL). Smooth muscle cells cultured on micropatterned and N-isopropylacrylamide-grafted (pNIPAm) polydimethylsiloxane (PDMS), a small portion of which was covered by aligned electrospun scaffolding, resulted in a single sheet of unidirectionally aligned cells. Herein we combine cell sheet technology and electrospun scaffolding to rapidly generate circumferentially aligned tubular constructs of human aortic smooth muscles cells with contractile gene expression for use as tissue engineered blood vessel media.