Method for in-vitro construction of vascular endodermis with flow shear stress resistant and platelet aggregation resistant functions

Abstract

The invention provides a method for in-vitro construction of vascular endodermis with flow shear stress resistant and platelet aggregation resistant functions. According to the method, flow shear stress increased gradually is slowly applied to vascular endothelial cells on the surface of an artificial blood vessel inner cavity so that the vascular endothelial cells gradually adapt to the shear force environment. The method comprises the following steps of connecting an artificial blood vessel with vascular endothelial cells growing on the surface of an inner cavity to a pipe of a shear stress inducible system, starting a liquid driving pump of the shear stress inducible system so that an initial shear force of 0-2dyne / cm<2> is applied to the vascular endothelial cells, adjusting the liquid driving pump every once in a while so that the shear stress on the vascular endothelial cells is improved by a certain ratio than shear stress in the previous stage until the shear stress on the vascular endothelial cells is equal to average or peak shear stress resisted by the arterial endothelial cells in a transplantation site, keeping the shear stress in a level of the average or peak shear stress resisted by the arterial endothelial cells in the transplantation site and carrying out endothelial cell culture for 24-72h.

Description

technical field [0001] The invention relates to the fields of cell culture and tissue engineering, and more specifically relates to a method for constructing a blood vessel endothelial layer in vitro with the functions of resisting flow shear force of physiological size and resisting platelet aggregation. Background technique [0002] The use of tissue engineering to construct small-caliber artificial vascular implants in vitro is an important way to solve the shortage of clinically available small-caliber vascular grafts. However, at present, small-caliber artificial blood vessels cannot be used clinically. An important reason is that acute thrombosis and long-term restenosis of the transplantation site are prone to occur after small-caliber artificial blood vessel transplantation. Lack of an intact, functional endothelial cell layer on the inner surface. [0003] The complete vascular endothelial cell layer in the body has the functions of anti-platelet aggregation, anti-...

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Problems solved by technology

Schneider et al. constructed a small-diameter artificial blood vessel with a complete endothelial cell layer by culturing vascular endothelial cells on the inner surface of the artificial blood vessel. Within hours, 15-85% of endothelial cells are washed away by blood flow (Schneider, P., Hanson, S. & Price, T.1988. Durability of confluent endothelial cell monolayers on small caliber vascular prostheses in vitro Surgery 103, 456-462. )

Ott et al. applied flow shear force to endothelial cells cultured on vascular scaffolds in vitro, first at 1-2dyn / cm 2 Induced under the action of flow shear force for 3 days, and then at 25dyn / cm 2 Induced under the action of flow shear force for 3 days, it was finally found that shear stress induction improved the shear stress tolerance of vascular endothelial cells (Ott, M.J., Olson, J.L. & Ballermann, B.J.1995. Chronic in vitro fow promotes ultrastructural differentiation of endothelial cells.Endothelium 3,21-30), but the induced shear stress applied by Ott et al. on endothelial cells is too severe, which may damage the normal function of endothelial cells

Inducing too severe changes in shear stress is a common problem in the current work to improve the shear stress tolerance of endothelial cells by means of shear stress induction, because vascular endothelial cells are very sensitive to changes in shear stress, Tsou et al. found that vascular endothelial cells Cells can feel 0.25dyn / cm 2 Changes in shear stress, thereby altering gene expression (Tsou, J.K. et al. 2008.Spatial regulation of inflammation by human aortic endothelial cells in a linear gradient of shear stress. Microcirculation 15, 311-323), so drastic changes in shear stress may damage the normal function of vascular endothelial cells

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