	US 11,898,167 B2
	Reversible stencils for fabricating micro-tissues
Nathaniel Huebsch, San Bruno, CA (US); Bruce Conklin, San Francisco, CA (US); Kevin E. Healy, Moraga, CA (US); and Peter Loskill, Stuttgart (DE)
Assigned to The J. David Gladstone Institutes, a testamentary trust established under the Will of J. David Gladstone, San Francisco, CA (US); and The Regents of the University of California, Oakland, CA (US)
Filed by The Regents of the University of California, Oakland, CA (US); and The J. David Gladstone Institutes, a testamentary trust established under the Will of J. David Gladstone, San Francisco, CA (US)
Filed on Oct. 26, 2020, as Appl. No. 17/079,720.
Application 17/079,720 is a continuation of application No. 15/329,035, granted, now 10,851,344, previously published as PCT/US2015/044232, filed on Aug. 7, 2015.
Claims priority of provisional application 62/034,210, filed on Aug. 7, 2014.
Prior Publication US 2021/0123023 A1, Apr. 29, 2021
Int. Cl. C12M 1/32 (2006.01); C12N 5/077 (2010.01); C12M 3/00 (2006.01); C12M 1/12 (2006.01)

CPC C12N 5/0657 (2013.01) [C12M 21/08 (2013.01); C12M 23/12 (2013.01); C12M 25/14 (2013.01); C12N 2533/30 (2013.01); C12N 2535/10 (2013.01)]

22 Claims

1. A device for confining mammalian cells, forcing cellular/tissue alignment and tissue self-assembly, comprising:

a cell adhesion substrate; and

a removable elastomeric stencil overlay;

wherein the elastomeric stencil has one or more cut-out patterned microwells comprising two or more circular, oval, rectangular, square, V-shaped, or triangular holes, each hole joined to an adjacent hole by a canal (shaft); and

wherein the cell adhesion substrate binds cells within the holes and the canals of the cut-out pattern; and

wherein at least one of the microwells has at least two square holes with a side length “L” of 250 μm to 1000 μm, and the canal joining the two square holes has a longitudinal length “X” of 250-1000 μm and a traverse width “Y” of 50-200 μm.