US 12,168,782 B2
Physiological biomimetic culture system for heart slices
Tamer Mohamed, Goshen, KY (US)
Assigned to University of Louisville Research Foundation, Louisville, KY (US)
Appl. No. 17/415,239
Filed by University of Louisville Research Foundation, Louisville, KY (US)
PCT Filed Dec. 19, 2019, PCT No. PCT/US2019/067343
§ 371(c)(1), (2) Date Jun. 17, 2021,
PCT Pub. No. WO2020/159642, PCT Pub. Date Aug. 6, 2020.
Claims priority of provisional application 62/798,572, filed on Jan. 30, 2019.
Prior Publication US 2022/0064601 A1, Mar. 3, 2022
Int. Cl. C12N 5/077 (2010.01); G01N 33/50 (2006.01); G09B 23/30 (2006.01)
CPC C12N 5/0657 (2013.01) [G01N 33/5088 (2013.01); G09B 23/306 (2013.01); C12N 2500/02 (2013.01); C12N 2501/115 (2013.01); C12N 2529/00 (2013.01); C12N 2533/30 (2013.01)] 12 Claims
 
1. A method for culturing a mammalian heart organotypic slice, comprising:
culturing the slice in a culture medium, wherein the culture medium comprises fetal bovine serum (FBS), vascular endothelial cell growth factor (VEGF), fibroblast growth factor (FGF), and insulin-transferrin-selenium (ITS) and wherein the culture medium does not include 2, 3-butanedione monoxime (BDM) or any fatty acids added in addition to fatty acids present in the FBS; and
applying electrical stimulation to the slice while the heart slice is in culture, wherein the electrical stimulation has a frequency of 0.5 to 2 Hz and wherein the electrical stimulation has a voltage of 5 to 15 V, and wherein a viability of the heart slice measured on day 6 of culturing is not statistically significantly different from a viability of the heart slice measured on day 1 of culturing as measured by MTT assay.