US 12,109,239 B2
Methods for production of human hemogenic endothelial cells from pluripotent stem cells and compositions thereof
Qiang Feng, Natick, MA (US); Shi-Jiang Lu, Shrewsbury, MA (US); and Robert P. Lanza, Clinton, MA (US)
Assigned to Astellas Institute for Regenerative Medicine, Westborough, MA (US)
Filed by Astellas Institute for Regenerative Medicine, Marlborough, MA (US)
Filed on Dec. 8, 2020, as Appl. No. 17/114,685.
Application 17/114,685 is a continuation of application No. 15/677,770, filed on Aug. 15, 2017, granted, now 10,894,065.
Application 15/677,770 is a continuation of application No. 14/138,008, filed on Dec. 21, 2013, granted, now 9,763,984.
Claims priority of provisional application 61/787,476, filed on Mar. 15, 2013.
Claims priority of provisional application 61/740,699, filed on Dec. 21, 2012.
Prior Publication US 2021/0161964 A1, Jun. 3, 2021
Int. Cl. C12N 5/078 (2010.01); A61K 35/19 (2015.01)
CPC A61K 35/19 (2013.01) [C12N 5/0644 (2013.01); C12N 2501/04 (2013.01); C12N 2501/20 (2013.01); C12N 2501/2306 (2013.01); C12N 2501/2309 (2013.01); C12N 2501/2311 (2013.01); C12N 2501/30 (2013.01); C12N 2501/40 (2013.01); C12N 2501/727 (2013.01); C12N 2501/734 (2013.01); C12N 2501/91 (2013.01); C12N 2506/11 (2013.01); Y02A 50/30 (2018.01)] 20 Claims
 
1. A method for producing human hemogenic endothelial (PVE-HE) cells comprising culturing human pluripotent stem cells in vitro on an extracellular matrix, under feeder free conditions, and in a differentiation induction medium (DIM) comprising BMP4, bFGF and VEGF for at least 4 days, wherein the at least 4 days comprise a first period of low oxygen conditions comprising 1% to 10% oxygen and a second period of normoxic conditions, thereby forming a cell population comprising hemogenic endothelial cells, wherein the human pluripotent stem cells are differentiated into the hemogenic endothelial cells without embryoid body formation.