US 11,840,708 B2
Isogenic blood-brain barrier model
Eric V. Shusta, Madison, WI (US); Scott G. Canfield, Middleton, WI (US); Clive N. Svendsen, Los Angeles, CA (US); Sean P. Palecek, Verona, WI (US); and Gad D. Vatine, Los Angeles, CA (US)
Assigned to WISCONSIN ALUMNI RESEARCH FOUNDATION, Madison, WI (US); and CEDARS-SINAI MEDICAL CENTER, Los Angeles, CA (US)
Appl. No. 16/091,450
Filed by Wisconsin Alumni Research Foundation, Madison, WI (US); and Cedars-Sinai Medical Center, Los Angeles, CA (US)
PCT Filed Apr. 4, 2017, PCT No. PCT/US2017/025935
§ 371(c)(1), (2) Date Oct. 4, 2018,
PCT Pub. No. WO2017/176747, PCT Pub. Date Oct. 12, 2017.
Claims priority of provisional application 62/318,424, filed on Apr. 5, 2016.
Prior Publication US 2019/0093084 A1, Mar. 28, 2019
Int. Cl. C12N 5/071 (2010.01); C12N 5/0793 (2010.01); C12N 5/079 (2010.01)
CPC C12N 5/0697 (2013.01) [C12N 5/069 (2013.01); C12N 5/0619 (2013.01); C12N 5/0622 (2013.01); C12N 2501/11 (2013.01); C12N 2501/115 (2013.01); C12N 2502/081 (2013.01); C12N 2502/086 (2013.01); C12N 2502/28 (2013.01); C12N 2506/45 (2013.01); C12N 2533/90 (2013.01)] 26 Claims
 
1. A method of inducing blood brain barrier properties in induced pluripotent stem cell (iPSC)-derived brain microvascular endothelial cells (BMECs) so as to generate an isogenic blood brain barrier model comprising the steps of:
(a) co-culturing iPSC-derived BMECs on a permeable membrane with a combination of iPSC-derived astrocytes and iPSC-derived neurons;
wherein
(i) the permeable membrane separates the BMECs from the astrocytes and neurons,
(ii) the neurons and astrocytes are derived from EZ-spheres differentiated from iPSCs,
(iii) blood brain barrier properties are induced in the BMECs to form the isogenic blood brain barrier model,
(iv) the iPSC-derived astrocytes and iPSC-derived neurons are isogenic to the iPSC-derived BMECs and all three cell types are derived from the same iPSC cell population,
(v) the ratio of neurons to astrocytes in the co-culture is 1:3, and
(vi) the trans-endothelial electrical resistance (TEER) of the isogenic blood brain barrier model is between 700-900 Ω×cm2 and is maintained for at least 5 days.