US 12,454,677 B2
Methods for the maturation of cardiomyocytes on amniotic fluid cell-derived ECM, cellular constructs, and uses for cardiotoxicity and proarrhythmic screening of drug compounds
Travis Block, San Antonio, TX (US); Edward S. Griffey, San Antonio, TX (US); and Todd Herron, New Hudson, MI (US)
Assigned to STEMBIOSYS, INC., San Antonio, TX (US)
Filed by STEMBIOSYS, INC., San Antonio, TX (US)
Filed on Nov. 19, 2021, as Appl. No. 17/531,402.
Application 17/531,402 is a continuation of application No. 16/797,945, filed on Feb. 21, 2020, granted, now 11,220,671.
Claims priority of provisional application 62/808,690, filed on Feb. 21, 2019.
Prior Publication US 2022/0119770 A1, Apr. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/077 (2010.01); G01N 33/50 (2006.01)
CPC C12N 5/0657 (2013.01) [G01N 33/5061 (2013.01); C12N 2503/02 (2013.01); C12N 2506/45 (2013.01); C12N 2533/52 (2013.01); C12N 2533/54 (2013.01); C12N 2533/90 (2013.01)] 19 Claims
OG exemplary drawing
 
1. A method for determining the cardiotoxicity or proarrhythmic effect of a drug compound in vitro, the method comprising:
(i) obtaining immature cardiomyocytes derived from human induced pluripotent stem cells, wherein the immature cardiomyocytes exhibit a single nucleus;
(ii) contacting the immature cardiomyocytes with an extracellular matrix obtained from culturing isolated cells from amniotic fluid obtained from a human at greater than 37 weeks of gestational age (AFC-ECM), wherein the AFC-ECM comprises laminin, collagen alpha-1 (XVIII), basement membrane-specific heparan sulfate proteoglycan core protein, agrin, vimentin, and collagen alpha-2 (IV), or isoforms thereof;
(iii) culturing the immature cardiomyocytes with the AFC-ECM in a culture media to form a layer of mature cardiomyocytes on the AFC-ECM, wherein the mature cardiomyocytes exhibit two nuclei; and
(iv) contacting the drug compound with mature cardiomyocytes and observing for a change in the electrophysiology of the mature cardiomyocytes to confirm whether the drug compound has a cardiotoxic or proarrhythmic effect on the mature cardiomyocytes.