Microfluidic device
The present invention relates to a microfluidic device ( 1 ), preferably for producing a three-dimensional cell culture, having at least one chamber ( 2 ), and a fluid channel ( 3 ) which flows through at least part of the chamber ( 2 ) in order to provide a fluid stream which flows through the chamber ( 2 ) preferably continuously, wherein the chamber ( 2 ) is connected to a loading opening ( 4 ) and via the loading opening ( 4 ) can be loaded with hydrogel up to a desired fill level, characterized in that the chamber ( 2 ) comprises a main chamber ( 5 ) and a secondary chamber ( 6 ) connected to the main chamber ( 5 ), wherein, when the chamber ( 2 ) is being loaded with hydrogel up to the desired fill level, the secondary chamber is at least partially filled with hydrogel backed up from the main chamber ( 5 ).
1 . A microfluidic device ( 1 ) for producing a three-dimensional cell culture, the microfluidic device comprising:
a chamber ( 2 ) comprising a main chamber portion ( 5 ) and a secondary chamber portion ( 6 ) connected to the main chamber portion ( 5 );
a fluid channel ( 3 ) extending at least partially through the chamber ( 2 ) for providing and configured to enable a fluid to flow through the chamber ( 2 ); and
a loading opening ( 4 ) configured to load hydrogel into the chamber ( 2 ), wherein the chamber ( 2 ) is connected to the loading opening ( 4 ) and is configured to be filled with hydrogel via the loading opening ( 4 ) up to a desired fill level,
wherein the main chamber portion ( 5 ) and the secondary chamber portion ( 6 ) are configured so that, when the chamber ( 2 ) is filled with hydrogel up to the desired fill level, both the main chamber portion ( 5 ) and the secondary chamber portion ( 6 ) are filled with hydrogel,
wherein the main chamber portion ( 5 ) has a main chamber portion bottom ( 8 ) and the secondary chamber portion ( 6 ) has a secondary chamber portion bottom ( 11 ),
wherein the main chamber portion bottom ( 8 ) is formed at a lower height level than the secondary chamber portion bottom ( 11 ),
wherein the secondary chamber portion ( 6 ) has a smaller volume than the main chamber portion ( 5 ).
2 . The microfluidic device ( 1 ) according to claim 1 , further comprising a connecting channel ( 7 ), wherein the main chamber portion ( 5 ) and the secondary chamber portion ( 6 ) are connected by the connecting channel ( 7 ), wherein the connecting channel ( 7 ) has a smaller cross section than the main chamber portion ( 5 ) and the secondary chamber portion ( 6 ).
3 . The microfluidic device ( 1 ) according to claim 1 , wherein the fluid channel ( 3 ) at least partially extends through the main chamber portion ( 5 ), wherein the fluid channel ( 3 ) comprises a fluid inlet ( 9 ) spaced apart from the main chamber portion bottom ( 8 ) in the main chamber portion ( 5 ) and a fluid outlet ( 10 ) spaced apart from the main chamber portion bottom.
4 . The microfluidic device ( 1 ) according to claim 3 , wherein the fluid channel ( 3 ) has a fluid channel bottom ( 16 ), wherein the fluid channel bottom ( 16 ) is formed at a higher height level than the main chamber portion bottom ( 8 ).
5 . The microfluidic device ( 1 ) according to claim 4 , wherein the fluid channel bottom ( 16 ) is formed at a lower height level than the secondary chamber bottom portion ( 11 ).
6 . The microfluidic device ( 1 ) according to claim 1 , wherein the loading opening ( 4 ) is connected to the secondary chamber portion ( 6 ).
7 . The microfluidic device ( 1 ) according to claim 1 , wherein the secondary chamber portion ( 6 ) is connected to the main chamber portion ( 5 ) on a side ( 12 ) of the main chamber portion ( 5 ) opposite the fluid channel ( 3 ).
8 . The microfluidic device ( 1 ) according to claim 1 , wherein the fluid channel ( 3 ) runs along a side wall ( 13 ) of the main chamber portion ( 5 ).
9 . The microfluidic device ( 1 ) according to claim 1 , further comprising a conduit ( 15 ) connecting the loading opening ( 4 ) to the chamber ( 2 ).
10 . The microfluidic device ( 1 ) according to claim 1 , wherein the chamber ( 2 ), the main chamber portion ( 5 ), the secondary chamber portion ( 6 ), the fluid channel ( 3 ), or a combination thereof comprises an antifouling surface coating.
11 . A carrier ( 17 ) comprising the microfluidic device ( 1 ) according to claim 1 .
12 . A method of producing a three-dimensional cell culture or an organoid using the microfluidic device ( 1 ) according to claim 1 , the method comprising loading the chamber ( 2 ) through the loading opening ( 4 ) with hydrogel up to the desired fill level.
13 . The method according to claim 12 , further comprising introducing cells into the chamber 2 through the loading opening 4 during the loading or after the loading or both during and after the loading.
14 . A method determining an influence of a chemical compound or of a physical parameter or of both on cells of a three-dimensional cell culture using the microfluidic device according to claim 1 , the method comprising:
a) detecting a first state of the cells;
b) contacting the cells with a chemical compound or changing a physical parameter within the microfluidic device or both;
c) detecting a second state of the cells; and
d) determining the influence of the chemical compound or of the physical parameter or of both on the cells by identifying a difference between the first state and the second state.
15 . The microfluidic device ( 1 ) according to claim 1 , wherein the fluid channel ( 3 ) is configured to enable a fluid to continuously pass through the chamber ( 2 ).
16 . The microfluidic device ( 1 ) according to claim 10 , wherein the antifouling surface coating is selected from the group consisting of a polyethylene glycol (PEG)-based polymer, a polysaccharide, agarose, a polyhydroxy polymer, poly(2-hydroxyethyl methacrylate) (poly-HEMA), 2-methacryloyloxyethyl phosphorylcholine (MPC), dextran, hydroxyethyl cellulose (HEC), a natural polymer, an S-layer protein, and combinations thereof.
17 . The method according to claim 13 , wherein the cells are selected from the group consisting of a stem cell, a primary cell, a fibroblast, a cartilage cell, an endothelial cell, an epithelial cell, an adipose cell, an induced pluripotent stem cells (IPS), an osteoclast, an osteoblast, and an osteocyte or a combination thereof.