US 12,241,085 B2
Human pluripotent stem cell derived neurodegenerative disease models on a microfluidic chip
Samuel Sances, Santa Monica, CA (US); Alexander Laperle, North Hollywood, CA (US); Nur Yucer, Los Angeles, CA (US); and Clive N. Svendsen, Pacific Palisades, CA (US)
Assigned to Cedars-Sinai Medical Center, Los Angeles, CA (US)
Appl. No. 17/041,672
Filed by CEDARS-SINAI MEDICAL CENTER, Los Angeles, CA (US)
PCT Filed Apr. 5, 2019, PCT No. PCT/US2019/026178
§ 371(c)(1), (2) Date Sep. 25, 2020,
PCT Pub. No. WO2019/195798, PCT Pub. Date Oct. 10, 2019.
Claims priority of provisional application 62/816,785, filed on Mar. 11, 2019.
Claims priority of provisional application 62/755,282, filed on Nov. 2, 2018.
Claims priority of provisional application 62/653,697, filed on Apr. 6, 2018.
Prior Publication US 2021/0130774 A1, May 6, 2021
Int. Cl. C12N 5/079 (2010.01); C12M 1/12 (2006.01); C12M 1/34 (2006.01); C12M 3/06 (2006.01); C12N 5/071 (2010.01); C12N 5/074 (2010.01); C12N 5/0793 (2010.01); G01N 33/50 (2006.01)
CPC C12N 5/0622 (2013.01) [C12M 23/16 (2013.01); C12M 25/02 (2013.01); C12M 41/46 (2013.01); C12N 5/0619 (2013.01); C12N 5/0696 (2013.01); C12N 5/0697 (2013.01); G01N 33/5058 (2013.01); C12N 2502/081 (2013.01); C12N 2502/086 (2013.01); C12N 2502/45 (2013.01); C12N 2506/45 (2013.01)] 9 Claims
OG exemplary drawing
 
1. A method of culturing cells, comprising:
providing
(i) astrocytes and brain microvascular endothelial cells (BMECs),
(ii) neurons,
(iii) microglia, and
(iv) a microfluidic device comprising a membrane comprising a top surface and a bottom surface;
seeding the BMECs on the bottom surface to create seeded endothelial cells and seeding the astrocytes on the top surface to create seeded astrocytes, the BMECs and astrocytes being seeded simultaneously;
seeding neurons on the top surface one or more days after seeding the BMECs and astrocytes to create seeded neurons;
seeding microglia on the top surface at one week to create seeded microglia; and
culturing the seeded endothelial cells, seeded astrocytes, seeded neurons and seeded microglia under flow conditions at a flow rate for a period of time.