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US 12,264,322 B2Granted
Minimal volume reprogramming of mononuclear cells
AssigneeFate Therapeutics, Inc.
InventorsPeter Flynn, Bahram Valamehr
Life of this patent
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Abstract
The invention provides compositions and methods for reprogramming minimal volumes of mononuclear cells. In particular aspects, the invention provides methods and compositions for reprogramming minimal volumes of umbilical cord blood obtained from cord blood segments from cryopreserved cord blood segments.
Claims23
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 1×106 of 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%.
Dependencies are inferred from the claim text.