US 12,258,585 B2
Methods of producing multi-layered tubular tissue constructs
Katharina Theresa Kroll, Somerville, MA (US); Kimberly A. Homan, Somerville, MA (US); Mark A. Skylar-Scott, Brookline, MA (US); Sebastien G. M. Uzel, Cambridge, MA (US); David B. Kolesky, Cambridge, MA (US); Patrick Lustenberger, Herrliberg (CH); and Jennifer A. Lewis, Cambridge, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Appl. No. 16/631,677
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
PCT Filed Jul. 20, 2018, PCT No. PCT/US2018/043042
§ 371(c)(1), (2) Date Jan. 16, 2020,
PCT Pub. No. WO2019/018737, PCT Pub. Date Jan. 24, 2019.
Claims priority of provisional application 62/535,557, filed on Jul. 21, 2017.
Prior Publication US 2020/0164109 A1, May 28, 2020
Int. Cl. C12N 5/071 (2010.01); A61F 2/06 (2013.01); A61L 27/28 (2006.01); A61L 27/36 (2006.01); A61L 27/38 (2006.01); A61L 27/50 (2006.01); B33Y 10/00 (2015.01); B33Y 70/00 (2020.01); B33Y 80/00 (2015.01); C12N 5/077 (2010.01); A61F 2/04 (2013.01)
CPC C12N 5/0691 (2013.01) [A61F 2/06 (2013.01); A61L 27/28 (2013.01); A61L 27/3625 (2013.01); A61L 27/3629 (2013.01); A61L 27/3633 (2013.01); A61L 27/3826 (2013.01); A61L 27/3882 (2013.01); A61L 27/3891 (2013.01); A61L 27/507 (2013.01); B33Y 10/00 (2014.12); B33Y 70/00 (2014.12); B33Y 80/00 (2014.12); C12N 5/0656 (2013.01); C12N 5/0661 (2013.01); C12N 5/069 (2013.01); C12N 5/0697 (2013.01); A61F 2002/041 (2013.01); A61F 2002/043 (2013.01); A61F 2002/044 (2013.01); A61F 2002/045 (2013.01); A61F 2002/046 (2013.01); A61F 2002/048 (2013.01); A61F 2002/065 (2013.01); A61F 2240/001 (2013.01); A61L 2420/08 (2013.01); C12N 2513/00 (2013.01); C12N 2533/54 (2013.01); C12N 2533/56 (2013.01); C12N 2533/90 (2013.01)] 25 Claims
 
1. A method of producing a perfusable multi-layered tubular tissue construct, comprising:
depositing on a substrate one or more cell-laden filaments, each cell-laden filament comprising:
a plurality of concentric and coaxial cell-laden ink layers, each cell-laden ink layer comprising one or more predetermined cell types and extending at least a portion of the length of the cell-laden filament, wherein the one or more predetermined cell types are cell aggregates or clusters of cells, and
a core comprising a fugitive ink, wherein the fugitive ink serves as a template for an open perfusable lumen within the cell-laden filament;
removing the fugitive ink to create the open perfusable lumen;
seeding the lumen with endothelial cells by:
providing the endothelial cells with the fugitive ink, wherein the endothelial cells remain in the open perfusable lumen after the fugitive ink is removed; and/or
injecting a suspension of endothelial cells into the open perfusable lumen after removing the fugitive ink; and
exposing the one or more cell-laden filaments to fluid perfusion to induce cell proliferation and development, thereby producing the perfusable multi-layered tubular tissue construct.