Microfluidic devices and methods incorporating assay units with multiple 3D scaffold regions
View Patent ↗A microfluidic device includes multiple microfluidic assay units arranged on a substrate, with each assay unit including multiple scaffold regions each containing a three-dimensional scaffold with associated cells, and a media channel surrounding a fluid-permeable boundary portion of at least a second scaffold region, wherein a fluid-permeable interface between the media channel and the second scaffold region comprises a curved shape spanning an arc of more than 90 degrees. A third scaffold region may be provided. Boundaries between different scaffold regions, and between a scaffold region and the media channel, may include microposts that may be spaced apart in a curved configuration. A method for performing an assay utilizing such a device is further provided.
1. A microfluidic device comprising:
a substrate; and
a plurality of microfluidic assay units arranged on the substrate, wherein each microfluidic assay unit of the plurality of microfluidic assay units comprises:
a first scaffold region containing a first three-dimensional scaffold comprising one or more first cells;
a second scaffold region containing a second three-dimensional scaffold comprising one or more second cells, wherein the second scaffold region surrounds, and is in contact with, a fluid-permeable boundary portion of the first scaffold region; and
a media channel surrounding a fluid-permeable boundary portion of the second scaffold region, and permitting fluid communication between contents of the media channel and at least the second scaffold region, wherein a fluid-permeable interface between the media channel and the second scaffold region comprises a curved shape spanning an arc of more than 90 degrees.
2. The microfluidic device of claim 1 , wherein:
each microfluidic assay unit further comprises:
a first inlet port in fluid communication with the first scaffold region;
a second inlet port and a second outlet port in fluid communication with the second scaffold region; and
a media channel inlet port and a media channel outlet port in fluid communication with the media channel; and
each microfluidic assay unit, including a respective first scaffold region, second scaffold region, media channel, first inlet port, second outlet port, media channel inlet port, and media channel outlet port, is arranged within a circular footprint.
3. The microfluidic device of claim 2 , wherein microfluidic assay units of the plurality of microfluidic assay units are arranged on the substrate in a two-dimensional array, with the circular footprint of each microfluidic assay unit being spaced apart from the circular footprint of each other microfluidic assay unit of the plurality of microfluidic assay units.
4. The microfluidic device of claim 1 , wherein:
for each microfluidic assay unit, the media channel comprises an inlet segment coupled with the media channel inlet port, an outlet segment coupled with the media channel outlet port, and a main segment arranged between the inlet segment and the outlet segment; and
the main segment comprises a greater width than each of the inlet segment and the outlet segment.
5. The microfluidic device of claim 4 , wherein the main segment contacts the fluid-permeable boundary portion of the second three-dimensional scaffold region.
6. The microfluidic device of claim 4 , wherein for each microfluidic assay unit, the main segment comprises a substantially constant width.
7. The microfluidic device of claim 5 , wherein for each microfluidic assay unit, the main segment comprises a semi-annular shape.
8. The microfluidic device of claim 1 , wherein each microfluidic assay unit further comprises:
a plurality of first microposts arranged between the first scaffold region and the second scaffold region; and
a plurality of second microposts arranged at a boundary of the second scaffold region, and arranged between the second scaffold region and the media channel.
9. The microfluidic device of claim 1 , wherein the first three-dimensional scaffold and the second three-dimensional scaffold each comprise a hydrogel.
10. The microfluidic device of claim 1 , wherein for each microfluidic assay unit, the fluid-permeable interface between the media channel and the second scaffold region comprises a curved shape spanning an arc of at least 135 degrees.
11. The microfluidic device of claim 1 , wherein for each microfluidic assay unit, the fluid-permeable interface between the media channel and the second scaffold region comprises a curved shape spanning an arc of at least 180 degrees.
12. The microfluidic device of claim 1 , wherein for each microfluidic assay unit:
the one or more first cells comprises at least one of migratory cells, stem cells, and tumor cells; and
the one or more second cells comprises at least one of: adipocytes, fibroblasts, macrophages, T-cells, and monocytes.
13. The microfluidic device of claim 1 , wherein:
each microfluidic assay unit further comprises a third scaffold region containing a third three-dimensional scaffold comprising one or more third cells, wherein the third scaffold region surrounds, and is in contact with, the fluid-permeable boundary portion of the second scaffold region; and
for each microfluidic assay unit, the third scaffold region is arranged between the second scaffold region and the media channel, wherein the fluid-permeable interface between the media channel and the second scaffold region includes a portion of the third scaffold region.
14. The microfluidic device of claim 13 , further comprising:
a plurality of first microposts arranged between the first scaffold region and the second scaffold region;
a plurality of second microposts arranged between the second scaffold region and the third scaffold region; and
a plurality of third microposts arranged between the third scaffold region and the media channel.
15. The microfluidic device of claim 13 , wherein for each microfluidic assay unit:
the one or more first cells comprises at least one of migratory cells, stem cells, and tumor cells;
the one or more second cells comprises at least one of adipocytes and fibroblasts; and
the one or more third cells comprises at least one of macrophages, T-cells, monocytes, vascular cells, endothelial cells, smooth muscle cells, and pericytes.
16. A method for performing an assay utilizing a microfluidic device that comprises a substrate and a plurality of microfluidic assay units arranged on the substrate, wherein each microfluidic assay unit comprises (i) a first scaffold region, (ii) a second scaffold region containing a second three-dimensional scaffold comprising one or more second cells, wherein the second scaffold region surrounds, and is in contact with, a fluid-permeable boundary portion of the first scaffold region, and (iii) a media channel surrounding a fluid-permeable boundary portion of the second scaffold region, wherein a fluid-permeable interface between the media channel and the second scaffold region comprises a curved shape spanning an arc of more than 90 degrees, the method comprising:
forming a first three-dimensional scaffold comprising a plurality of first cells in the first scaffold region;
forming a second three-dimensional scaffold comprising a plurality of second cells in the second scaffold region; and
supplying liquid media to the media channel to permit fluid communication between contents of the media channel and at least the second scaffold region.
17. The method of claim 16 , further comprising, for each microfluidic assay unit, extracting liquid media from the media channel, and analyzing the extracted liquid media.
18. The method of claim 16 , further comprising, for each microfluidic assay unit:
digesting at least a portion of the first three-dimensional scaffold in the first scaffold region;
removing at least some first cells of the plurality of first cells from the digested first three-dimensional scaffold of the first scaffold region; and
analyzing the at least some first cells.
19. The method of claim 16 , further comprising, for each microfluidic assay unit:
digesting at least a portion of the second three-dimensional scaffold in the second scaffold region;
removing at least some second cells of the plurality of second cells from the digested second three-dimensional scaffold of the second scaffold region; and
analyzing the at least some second cells.
20. The method of claim 16 , wherein:
each microfluidic assay unit further comprises (iv) a third scaffold region that surrounds, and is in contact with, the fluid-permeable boundary portion of the second scaffold region, with the third scaffold region arranged between the second scaffold region and the media channel, and with the fluid-permeable interface between the media channel and the second scaffold region including a portion of the third scaffold region;
the method further comprises forming a third three-dimensional scaffold comprising a plurality of third cells in the third scaffold region; and
the supplying of liquid media to the media channel permits fluid communication between contents of the media channel and at least the second scaffold region through the third scaffold region.
21. The method of claim 20 , further comprising, for each microfluidic assay unit:
digesting at least a portion of the third three-dimensional scaffold in the third scaffold region;
removing at least some third cells of the plurality of third cells from the digested third three-dimensional scaffold of the third scaffold region; and
analyzing the at least some third cells.
22. The method of claim 16 , further comprising performing imaging analysis of at least a portion of each microfluidic assay unit.