Culture vessels containing 3D cell culture substrates with diffusion structures
A cell culture device comprises a multi-well cell culture plate comprising a plurality of wells, each well comprising a top, a bottom, and a sidewall disposed between the top and the bottom and having an interior surface comprising an ultra-low attachment surface. A plurality of scaffolds are disposed within wells of the multi-well cell culture plate, each scaffold comprising a cell-adherent surface. In some embodiments, the scaffold comprises a fiber scaffold. In some embodiments, the scaffold comprises an artificial vascular scaffold. In some embodiments, cell culture devices comprise a plurality of hydrogel scaffolds disposed in a multi-well cell culture plate, the plurality of hydrogel scaffolds comprising hydrogel fibers of differing lengths, wherein opposite ends of a hydrogel fiber are disposed in different wells within the multi-well cell culture plate to create interconnected wells.
1 . A cell culture device comprising: a multi-well cell culture plate comprising a plurality of wells, each well comprising a top aperture, a bottom, and a sidewall disposed between the top and the bottom, wherein each well is configured with an interior surface comprising a cell non-adherent surface; and a plurality of scaffolds disposed within the plurality of wells of the multi-well cell culture plate, wherein at least one scaffold is disposed in each well and each scaffold comprising a cell-adherent surface; wherein the bottom comprises a plurality of microcavities; and wherein a scaffold is anchored to a bottom portion of each microcavity of the plurality of microcavities.
2 . The device of claim 1 , wherein the bottom comprises a hemispherical shape.
3 . The device of claim 1 , wherein at least one end of the scaffold is anchored to a bottom portion of each well.
4 . The device of claim 1 , wherein the scaffold has an average length in a range of 100 pm to 1000 pm.
5 . The device of claim 1 , wherein the scaffold has an average width in a range of 10 pm to 100 pm.
6 . The device of claim 1 , wherein a scaffold of the plurality of scaffolds comprises a fiber scaffold.
7 . The device of claim 6 , wherein the fiber scaffold is formed from polyvinylalcohol, polyacrylamide, polyvinylpyrrolidone, poly(2-hydroxyethyl methacrylate), polystyrene, polypropylene, polygalacturonic acid, and/or combinations thereof.
8 . The device of claim 7 , wherein the fiber scaffold comprises a plurality of fibers.
9 . The device of claim 8 , wherein each fiber in the plurality of fibers is anchored to a bottom portion of each well.
10 . The device of claim 9 , wherein anchored ends of individual fibers are distanced 100 μm to 200 μm apart from one another in each well.
11 . The device of claim 1 , wherein a scaffold of the plurality of scaffolds comprises an artificial vascular scaffold.
12 . The device of claim 11 , wherein the artificial vascular scaffold comprises a hollow fiber.
13 . The device of claim 11 , wherein the artificial vascular scaffold comprises a plurality of hollow fibers.
14 . The device of claim 13 , wherein individual hollow fibers are anchored to the bottom of a well 100 μm to 200 μm from one another.
15 . The device of claim 12 , wherein the hollow fiber is formed from a non-ionic polymer.
16 . The device of claim 15 , wherein the non-ionic polymer comprises polyvinylalcohol, polyacrylamide, polyvinylpyrrolidone, poly(2-hydroxyethyl methacrylate), polystyrene, polypropylene, or polygalacturonic acid.
17 . The device of claim 12 , wherein the hollow fiber further comprises a hydrogel coating.
18 . The device of claim 11 , wherein the artificial vascular scaffold comprises a hydrogel.
19 . A method of culturing three-dimensional (3D) cell cultures, the method comprising:
seeding cells in a cell culture device according to claim 1 , wherein the cells attach to the cell adherent surface of each scaffold disposed within the cell culture device; and
culturing the cells into a 3D cell culture by adding cell culture medium to the cell culture device to provide nutrients and oxygen, wherein the cells remain attached to each scaffold during addition or exchange of cell culture medium; and
optionally digesting each of the plurality of scaffolds.
20 . The method of claim 19 , further comprising imaging 3D cell cultures attached to each of the plurality of scaffolds of the cell culture device.