	US 12,264,322 B2
	Minimal volume reprogramming of mononuclear cells
Peter Flynn, San Diego, CA (US); and Bahram Valamehr, San Diego, CA (US)
Assigned to FATE THERAPEUTICS, INC., San Diego, CA (US)
Filed by Fate Therapeutics, Inc., San Diego, CA (US)
Filed on Feb. 23, 2021, as Appl. No. 17/183,263.
Application 16/007,928 is a division of application No. 14/731,403, filed on Jun. 4, 2015, granted, now 10,023,879, issued on Jul. 17, 2018.
Application 17/183,263 is a continuation of application No. 16/007,928, filed on Jun. 13, 2018, granted, now 10,954,529.
Claims priority of provisional application 62/007,924, filed on Jun. 4, 2014.
Prior Publication US 2021/0198687 A1, Jul. 1, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 48/00 (2006.01); A61K 35/545 (2015.01); C12N 15/85 (2006.01); A61K 35/12 (2015.01); A61K 35/51 (2015.01)

CPC C12N 15/85 (2013.01) [A61K 35/545 (2013.01); A61K 2035/124 (2013.01); A61K 35/51 (2013.01); C12N 2740/16043 (2013.01); C12N 2800/108 (2013.01); C12N 2800/30 (2013.01); C12N 2840/20 (2013.01)]

23 Claims

1. A method for producing induced pluripotent stem cells (iPSC) from an individual segment of an umbilical cord blood unit (UCB) comprising:

providing an individual segment of UCB comprising about 100 to about 1×10⁶of CD34+ mononuclear cells, wherein said CD34+ mononuclear cells comprise CD45+/CD34+/Lineage− cells, and wherein the individual segment of UCB has a minimal volume of less than 500 μL; and

reprogramming the CD34+ mononuclear cells to iPSC using a non-integrating method, wherein the CD34+ mononuclear cells comprise at least one exogenous polypeptide selected from the group consisting of an octamer-binding transcription factor 4 (OCT4) polypeptide, a sex determining region Y box 2 (SOX2) polypeptide and a Simian virus 40 large antigen (SV40LT) polypeptide, wherein reprogrammed iPSC co-express SSEA4 and TRA181 with a reprograming efficiency of at least 5%.