Methods, substrates, and systems useful for cell seeding of medical grafts
View Patent ↗Described are methods, cell growth substrates, and devices that are useful in preparing cell-containing graft materials for administration to patients. Tubular passages can be defined in cell growth substrates to promote distribution of cells into the substrates. Also described are methods and devices for preparing cell-seeded graft compositions, methods and devices for preconditioning cell growth substrates prior to application of cells, and cell seeded grafts having novel substrates, and uses thereof.
1. A cell growth substrate, comprising: a cell growth substrate for supporting the growth of cells, the cell growth substrate including at least one elongate tubular passage, the elongate tubular passage having passage walls defining a lumen extending from a first lumen opening at a surface of the cell growth substrate into an interior region of the cell growth substrate, with said lumen configured to receive flow of a cell-containing liquid medium to distribute cells into the interior region of the cell growth substrate; wherein the cell growth substrate includes a cell growth substrate body comprised of a porous matrix material suitable for supporting cellular growth, with the cell growth substrate body including a first decellularized extracellular matrix tissue sheet and a second decellularized extracellular matrix tissue sheet, the first decellularized extracellular matrix sheet connected to the second decellularized extracellular matrix tissue sheet in such a fashion as to define the at least one elongate tubular passage between the first sheet and the second sheet, the passage walls of the elongate tubular passage having (i) a first passage wall portion defined by a face of the first decellularized extracellular matrix tissue sheet, (i) a second passage wall portion defined by a face of the second decellularized extracellular matrix tissue sheet, (W) a first seam defined by a first connection between the face of the first decellularized extracellular matrix tissue sheet and the face of the second decellularized extracellular matrix sheet, and (iv) a second seam defined by a second connection between the face of the first decellularized extracellular matrix tissue sheet and the face of the second decellularized extracellular matrix sheet; and the first decellularized extracellular matrix tissue sheet and the second decellularized extracellular matrix tissue sheet being connected to one another along a connected region surrounding the at least one elongate tubular passage.
2. The cell growth substrate of claim 1 , also comprising a synthetic polymeric tubular element exterior of the substrate and fluidly coupled to the lumen opening.
3. The cell growth substrate of claim 1 , comprising a plurality of said tubular passages.
4. The cell growth substrate of claim 1 , wherein the at least one elongate tubular passage includes at least one primary tubular passage with, at least one secondary tubular passage branching from the primary tubular passage.
5. The cell growth substrate of claim 1 wherein the tubular passage extends from the first lumen opening to a second lumen opening spaced on said substrate from the first lumen opening.
6. The cell growth substrate of claim 1 , wherein the tubular passage terminates within the substrate.
7. The cell growth substrate of claim 1 , wherein the first decellularized extracellular matrix tissue and second decellularized extracellular matrix tissue sheets are remodelable collagenous extracellular matrix sheet materials.
8. The cell growth substrate of claim 1 , wherein the first decellularized extracellular matrix tissue sheet and the second decellularized extracellular matrix tissue sheet retain growth, factors, glycosaminoglycans, and proteoglycans from an animal source tissue.
9. The cell growth substrate of claim 1 , wherein the at least one elongate tubular passage comprises a tortuous tubular passage.
10. The cell growth substrate of claim 9 , wherein the tortuous tubular passage includes a first plurality of bends occurring opposite a second plurality of bends, with bends of the first plurality of bends and the second plurality of bends situated in opposed directions.
11. The cell growth substrate of claim 1 , wherein the first decellularized extracellular matrix tissue sheet is bonded, fused, glued, or crosslinked to the second decellularized extracellular matrix tissue sheet in the connected region.
12. The cell growth substrate of claim 1 , comprising a plurality of said tubular passages, and further wherein the tubular passages are separated from one another by intervening connected regions in which the face of the first decellularized extracellular matrix tissue sheet is connected to the face of the second decellularized extracellular matrix tissue sheet.
13. The cell growth substrate of claim 12 , comprising tubular elements comprising a persistent or bioresorbable polymer, the tubular elements received within the tubular passages.
14. The cell growth substrate of claim 13 , wherein the tubular elements include walls that are porous or that have holes or perforations.
15. The cell growth substrate of claim 1 , wherein the at least one elongate tubular passage includes a common feed passage fluidly coupled to a plurality of passages.
16. The cell growth substrate of claim 1 , further comprising a cellular composition in the interior region of the cell growth substrate.
17. A method for preparing a cell-seeded material, comprising:
providing the cell growth substrate of claim 1 ;
connecting a cell growth substrate to a source of a liquid suspension of cells, the connecting including fluidly associating the elongate passage with a transport lumen of the source; and
transporting amounts of the liquid suspension of cells through the transport lumen and into the elongate passage so as to deliver cells to the interior region of the substrate.
18. The method of claim 17 , wherein said connecting includes inserting the growth substrate into a cell-seeding chamber having an input tube fluidly connected to the source of liquid suspension of cells, and fluidly coupling the input tube to the first lumen opening of the substrate.