Bioengineered Allogeneic Valve
The present disclosure relates to methods for recellularization of valves in valve-bearing veins. This method is useful for producing an allogeneic venous valve, wherein a donor valve-bearing vein is decellularized and then recelluiarized using whole blood or bone marrow stem cells. The allogeneic valves produced by the methods disclosed herein are advantageous for implantation, transplantation, or grafting into patients with vascular diseases.
1 . A method of recellularization of a valve in a vein, comprising introducing blood comprising progenitor cells for endothelial cells and progenitor cells for smooth muscle cells to the lumen of a decellularized vein, and culturing the cells in the lumen of the decellularized vein, thereby recellularizing the valve in the vein.
2 . The method of claim 1 , wherein the blood is peripheral venous blood or whole blood.
3 . The method of claim 2 , wherein the peripheral venous blood or the whole blood is introduced to the decellularized vein by injection or perfusion.
4 . The method of claim 3 , further comprising culturing the cells by perfusion of endothelial cell medium and smooth muscle cell medium.
5 . The method of claim 4 , wherein the perfusion of the endothelial cell medium and the smooth muscle cell medium are in alternation.
6 . The method of one of the above claims, wherein the recellularized valve is CD31 positive, vWF positive, smooth muscle actin positive, and has nuclei.
7 . The method of one of the above claims, wherein the recellularized valve has mechanical properties of withstanding force at first peak at or above 0.8 N.
8 . The method of one of the above claims, wherein the recellularized valve has a closure time of equal to or less than 0.5 seconds.
9 . A method of treating chronic venous insufficiency (CVI), deep vein thrombosis (DVT), and/or leg ulceration in a subject in need thereof, comprising introducing a recellularized valve-bearing segment of a vein to the subject, wherein the valve is recellularized by a method comprising:
a) decellularizing a valve-bearing segment of a vein, wherein the vein is allogeneic to the subject;
b) collecting blood from the subject, wherein the blood comprises progenitor cells for endothelial cells and progenitor cells for smooth muscle cells;
c) perfusing the decellularized valve-bearing with the collected blood;
d) culturing the cells in the lumen of the decellularized valve-bearing vein; thereby recellularizing the decellularized valve of the vein; and
e) grafting the recellularized valve-bearing vein to the subject; wherein the grafting treats CVI and/or leg ulceration in the subject.
10 . The method of claim 9 , wherein the leg ulcer is recurrent and is due to deep venous reflux and/or venous hypertension.
11 . The method of claim 10 , wherein the blood is peripheral venous blood or whole blood.
12 . The method of claim 11 , wherein the peripheral venous blood or the whole blood is introduced to the decellularized vein by injection or perfusion.
13 . The method of claim 12 , further comprising culturing the cells by perfusion of endothelial cell medium and smooth muscle cell medium.
14 . The method of claim 13 , wherein the perfusion of the endothelial cell medium and the smooth muscle cell medium are in alternation.
15 . The method of claims 9 - 14 , wherein the recellularized valve is CD31 positive, vWF positive, smooth muscle actin positive, and has nuclei.
16 . The method of claims 9 - 14 , wherein the recellularized valve has mechanical properties of withstanding force at first peak at or above 0.8 N.
17 . The method of claims 9 - 14 , wherein the recellularized valve has a closure time of equal to or less than 0.5 seconds.
18 . A method of recellularization of a valve in a valve-bearing vein, wherein the recellularized valve is a functional valve, the method comprising:
a) harvesting a segment of a vein from a subject in need of the recellularized valve in a vein;
b) decellularizing the valve-bearing vein;
c) collecting blood from the subject, wherein the blood comprises progenitor cells for endothelial cells and progenitor cells for smooth muscle cells;
d) perfusing the decellularized valve-bearing vein with the collected blood; and
e) culturing the cells in the decellularized valve-bearing vein, thereby recellularizing the decellularized valve in the vein.
19 . The method of claim 18 , wherein the recellularization restores competence and tolerance of reflux pressure of the recellularized valve.
20 . The method of claim 18 , wherein the blood is peripheral venous blood or whole blood.
21 . The method of claim 20 , wherein the peripheral venous blood or the whole blood is introduced to the decellularized vein by injection or perfusion.
22 . The method of claim 21 , further comprising culturing the cells by perfusion of endothelial cell medium and smooth muscle cell medium.
23 . The method of claim 22 , wherein the perfusion of the endothelial cell medium and the smooth muscle cell medium are in alternation.
24 . The method of claims 18 - 23 , wherein the recellularized valve is CD31 positive, vWF positive, smooth muscle actin positive, and has nuclei.
25 . The method of claims 18 - 24 , wherein the recellularized valve has mechanical properties of withstanding force at first peak at or above 0.8 N.
26 . The method of claims 18 - 24 , wherein the recellularized valve has a closure time of equal to or less than 0.5 seconds.