	US 12,139,724 B2
	Methods of making red blood cells and platelets in vitro and uses thereof
Tara L. Deans, Cottonwood Heights, UT (US)
Assigned to University of Utah Research Foundation, Salt Lake City, UT (US)
Filed by University of Utah Research Foundation, Salt Lake City, UT (US)
Filed on Apr. 15, 2022, as Appl. No. 17/721,690.
Application 17/721,690 is a continuation of application No. 15/745,305, granted, now 11,319,528, previously published as PCT/US2016/042084, filed on Jul. 13, 2016.
Claims priority of provisional application 62/231,690, filed on Jul. 13, 2015.
Prior Publication US 2022/0251510 A1, Aug. 11, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/078 (2010.01); C12N 5/074 (2010.01); C12N 5/0789 (2010.01); C12N 15/85 (2006.01)

CPC C12N 5/0644 (2013.01) [C12N 5/0641 (2013.01); C12N 5/0647 (2013.01); C12N 15/85 (2013.01); C12N 5/0696 (2013.01); C12N 2501/14 (2013.01); C12N 2501/145 (2013.01); C12N 2510/00 (2013.01)]

14 Claims

1. A method of producing a platelet or red blood cell comprising a therapeutic peptide, the method comprising:

a) providing a feeder cell;

b) providing a genetically engineered fed cell, wherein the genetically engineered fed cell comprises

i) a first genetic circuit, wherein the first genetic circuit comprises GATA-1 and/or HoxB4, one or more recombination sites, one or more recombinases, and one or more promoters operably linked to the GATA-1 and/or HoxB4, one or more recombination sites and one or more recombinases; and

ii) a second genetic circuit, wherein the second genetic circuit comprises a gene of interest capable of encoding a therapeutic peptide, one or more recombination sites, one or more recombinases, and one or more promoters operably linked to the gene of interest, one or more recombination sites and one or more recombinases;

c) culturing the feeder cell in a) with the genetically engineered fed cell in b) in a media under conditions that permit the genetically engineered fed cell to differentiate into platelet progenitor stem cells and/or differentiate into red blood progenitor stem cells and to express the gene of interest; and

d) producing the platelet progenitor stem cells and/or red blood progenitor stem cells comprising the therapeutic peptide encoded by the gene of interest.