US 12,378,519 B2
Microfluidic model of the blood brain barrier
S. Jordan Kerns, Reading, MA (US); Norman Wen, West Roxbury, MA (US); Carolina Lucchesi, Westwood, MA (US); Christopher David Hinojosa, Cambridge, MA (US); Jacob Fraser, Somerville, MA (US); Geraldine Hamilton, Boston, MA (US); Gad Vatine, Los Angeles, CA (US); Samuel Sances, Santa Monica, CA (US); Clive Svendsen, Pacific Palisades, CA (US); Daniel Levner, Brookline, MA (US); and Dhruv Sareen, Porter Ranch, CA (US)
Assigned to EMULATE, INC., Boston, MA (US); and CEDARS-SINAI MEDICAL CENTER, Los Angeles, CA (US)
Filed by EMULATE, Inc., Boston, MA (US); and Cedars-Sinai Medical Center, Los Angeles, CA (US)
Filed on Dec. 5, 2023, as Appl. No. 18/529,198.
Application 18/529,198 is a continuation of application No. 15/768,736, previously published as PCT/US2016/057724, filed on Oct. 19, 2016.
Claims priority of provisional application 62/380,780, filed on Aug. 29, 2016.
Claims priority of provisional application 62/332,727, filed on May 6, 2016.
Claims priority of provisional application 62/277,723, filed on Jan. 12, 2016.
Claims priority of provisional application 62/243,642, filed on Oct. 19, 2015.
Prior Publication US 2024/0228954 A1, Jul. 11, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0793 (2010.01); C12M 1/12 (2006.01); C12N 5/071 (2010.01); C12N 5/079 (2010.01); C12Q 1/02 (2006.01); B01L 3/00 (2006.01); C12M 1/42 (2006.01); C12M 3/06 (2006.01)
CPC C12N 5/0619 (2013.01) [C12N 5/0622 (2013.01); C12N 5/069 (2013.01); C12N 5/0692 (2013.01); C12Q 1/02 (2013.01); B01L 3/5027 (2013.01); B01L 2200/0647 (2013.01); B01L 2300/0861 (2013.01); C12M 23/16 (2013.01); C12M 25/02 (2013.01); C12M 35/08 (2013.01); C12N 5/0618 (2013.01); C12N 2502/081 (2013.01); C12N 2502/086 (2013.01); C12N 2502/28 (2013.01); C12N 2506/45 (2013.01); C12N 2531/00 (2013.01); C12N 2533/52 (2013.01); C12N 2533/54 (2013.01); C12N 2533/56 (2013.01); C12N 2535/10 (2013.01); C12N 2539/00 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method of culturing cells, comprising:
a) providing a microfluidic device comprising a porous membrane, said membrane comprising a top surface and a bottom surface;
b) seeding brain microvascular endothelial cells on said bottom surface so as to create seeded-cells on said bottom surface of said membrane;
c) seeding stem cell-derived neuronal cells or neuronal cells differentiated from stem cells on said top surface to create seeded cells on said top surface of said membrane; and
d) culturing said top surface seeded cells under flow conditions that support the maturation of neurons and culturing said bottom surface seeded cells under conditions that support maturation of said brain microvascular endothelial cells wherein said neurons exhibit a more mature electrophysiology, in terms of one or more features selected from the group consisting of action potential patterns, negative sodium channel current, positive potassium channel current and action potential spikes of amplitude, duration and frequency, or increased frequency after exposure to said flow as compared to the same neurons cultured in a static culture.