US 11,718,823 B2
Methods and materials for rapid preparation of 3D spheroids/organoids
Bumsoo Han, West Lafayette, IN (US); George Tsu-Chih Chiu, West Lafayette, IN (US); and Cih Cheng, West Lafayette, IN (US)
Assigned to Purdue Research Foundation, West Lafayette, IN (US)
Filed by Purdue Research Foundation, West Lafayette, IN (US)
Filed on Dec. 1, 2020, as Appl. No. 17/108,009.
Claims priority of provisional application 62/969,177, filed on Feb. 3, 2020.
Prior Publication US 2021/0238541 A1, Aug. 5, 2021
Int. Cl. B33Y 10/00 (2015.01); B33Y 70/00 (2020.01); C12N 5/00 (2006.01); C12N 5/0775 (2010.01); C12N 5/071 (2010.01); C12N 5/09 (2010.01)
CPC C12N 5/0062 (2013.01) [B33Y 10/00 (2014.12); B33Y 70/00 (2014.12); C12N 5/0663 (2013.01); C12N 5/0693 (2013.01); C12N 5/0697 (2013.01); C12N 2513/00 (2013.01)] 6 Claims
 
1. A method of printing a cellular spheroid with a shell and a core, wherein the method comprises:
providing a first solution comprising a first hydrogel, a first collagen, and a plurality of first cells, wherein the first cells are configured to substantially stay in the shell of said cellular spheroid;
providing a second solution comprising a second hydrogel, a second collagen, and a plurality of second cells; wherein the second cells are configured to substantially stay in the core of said cellular spheroid;
wherein the first hydrogel and the second hydrogel may be same or different, wherein the first collagen and the second collagen may be same or different, wherein the first cells and the second cells are different, wherein the second cells have higher contractility than the first cells;
printing a plurality of droplets of the first solution on a substrate and incubating said first solution for gelation to form a first line array;
printing a plurality of droplets of the second solution on said substrate adjacent to the first line array and incubating said second solution for gelation to form a second line array, wherein the printing and the gelation of the second solution is carried out after the printing and the gelation of the first printed solution, wherein the incubated first solution first line array is adjacent to the incubated second solution second line array, thereby forming a cell-containing construct;
adding a cell culture medium to the cell-containing construct incubated first solution and the incubated second solution to allow the cell-containing construct incubated first solution and the incubated second solution to detach from the substrate and to shrink towards the direction of the second cells, which forms thereby forming a cellular spheroid with a shell and a core, wherein the shell comprises the first cells, and the core comprises the second cells; and
growing the cellular spheroid to a desired size, wherein the shell comprises the first cells, and the core comprises the second cells.