US 12,234,476 B2
VCN enhancer compositions and methods of using the same
Melissa Bonner, Natick, MA (US); and Olivier Negre, Cambridge, MA (US)
Assigned to BLUEBIRD BIO, INC., Somerville, MA (US)
Appl. No. 16/076,946
Filed by bluebird bio, Inc., Cambridge, MA (US)
PCT Filed Feb. 10, 2017, PCT No. PCT/US2017/017372
§ 371(c)(1), (2) Date Aug. 9, 2018,
PCT Pub. No. WO2017/139576, PCT Pub. Date Aug. 17, 2017.
Claims priority of provisional application 62/294,615, filed on Feb. 12, 2016.
Claims priority of provisional application 62/313,571, filed on Mar. 25, 2016.
Claims priority of provisional application 62/417,085, filed on Nov. 3, 2016.
Claims priority of provisional application 62/429,514, filed on Dec. 2, 2016.
Prior Publication US 2019/0284533 A1, Sep. 19, 2019
Int. Cl. C12N 15/867 (2006.01); C12N 5/0789 (2010.01); C12N 7/00 (2006.01)
CPC C12N 15/867 (2013.01) [C12N 5/0647 (2013.01); C12N 7/00 (2013.01); C12N 2501/02 (2013.01); C12N 2501/999 (2013.01); C12N 2510/00 (2013.01); C12N 2740/15011 (2013.01); C12N 2740/15041 (2013.01); C12N 2740/16011 (2013.01); C12N 2740/16041 (2013.01); C12N 2740/16043 (2013.01)] 17 Claims
 
1. A method of increasing the vector copy number (VCN) in a population of CD34+ hematopoietic stem or progenitor cells comprising culturing the cells for at least two hours in the presence of: (1) a lentiviral vector at a multiplicity of infection (MOI) of about 10 to about 30; (2) a prostaglandin E2(PGE2), or 16,16-dimethyl PGE2 ; and (3) poloxamer 338, wherein the increase of VCN achieved by culturing the cells for at least two hours in the presence of the lentiviral vector, poloxamer 338 and one of PGE2 or 16,16-dimethyl PGE2 is further enhanced beyond the increased VCN achieved by culturing the cells for at least two hours in the presence of the lentiviral vector and one of PGE2 or 16,16-dimethyl PGE2 and the absence of poloxamer 338.